Methods and Compositions for Diagnosis and Prognosis of Renal Injury and Renal Failure

ABSTRACT

The present invention relates to methods and compositions for monitoring, diagnosis, prognosis, and determination of treatment regimens in subjects suffering from or suspected of having a renal injury. In particular, the invention relates to using assays that detect one or more of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24 as diagnostic and prognostic biomarker assays in renal injuries.

The present application claims the benefit of U.S. Provisional Patent Application 62/455,503 filed Feb. 6, 2017, which is hereby incorporated by reference in its entirety including all tables, FIGURES and claims.

BACKGROUND OF THE INVENTION

The following discussion of the background of the invention is merely provided to aid the reader in understanding the invention and is not admitted to describe or constitute prior art to the present invention.

The kidney is responsible for water and solute excretion from the body. Its functions include maintenance of acid-base balance, regulation of electrolyte concentrations, control of blood volume, and regulation of blood pressure. As such, loss of kidney function through injury and/or disease results in substantial morbidity and mortality. A detailed discussion of renal injuries is provided in Harrison's Principles of Internal Medicine, 17^(th) Ed., McGraw Hill, New York, pages 1741-1830, which are hereby incorporated by reference in their entirety. Renal disease and/or injury may be acute or chronic. Acute and chronic kidney disease are described as follows (from Current Medical Diagnosis & Treatment 2008, 47^(th) Ed, McGraw Hill, New York, pages 785-815, which are hereby incorporated by reference in their entirety): “Acute renal failure is worsening of renal function over hours to days, resulting in the retention of nitrogenous wastes (such as urea nitrogen) and creatinine in the blood. Retention of these substances is called azotemia. Chronic renal failure (chronic kidney disease) results from an abnormal loss of renal function over months to years”.

Acute renal failure (ARF, also known as acute kidney injury, or AKI) is an abrupt (typically detected within about 48 hours to 1 week) reduction in glomerular filtration. This loss of filtration capacity results in retention of nitrogenous (urea and creatinine) and non-nitrogenous waste products that are normally excreted by the kidney, a reduction in urine output, or both. It is reported that ARF complicates about 5% of hospital admissions, 4-15% of cardiopulmonary bypass surgeries, and up to 30% of intensive care admissions. ARF may be categorized as prerenal, intrinsic renal, or postrenal in causation. Intrinsic renal disease can be further divided into glomerular, tubular, interstitial, and vascular abnormalities. Major causes of ARF are described in the following table, which is adapted from the Merck Manual, 17^(th) ed., Chapter 222, and which is hereby incorporated by reference in their entirety:

Type Risk Factors Prerenal ECF volume depletion Excessive diuresis, hemorrhage, GI losses, loss of intravascular fluid into the extravascular space (due to ascites, peritonitis, pancreatitis, or burns), loss of skin and mucus membranes, renal salt- and water-wasting states Low cardiac output Cardiomyopathy, MI, cardiac tamponade, pulmonary embolism, pulmonary hypertension, positive-pressure mechanical ventilation Low systemic vascular Septic shock, liver failure, antihypertensive drugs resistance Increased renal vascular NSAIDs, cyclosporines, tacrolimus, hypercalcemia, resistance anaphylaxis, anesthetics, renal artery obstruction, renal vein thrombosis, sepsis, hepatorenal syndrome Decreased efferent ACE inhibitors or angiotensin II receptor blockers arteriolar tone (leading to decreased GFR from reduced glomerular transcapillary pressure, especially in patients with bilateral renal artery stenosis) Intrinsic Renal Acute tubular injury Ischemia (prolonged or severe prerenal state): surgery, hemorrhage, arterial or venous obstruction; Toxins: NSAIDs, cyclosporines, tacrolimus, aminoglycosides, foscarnet, ethylene glycol, hemoglobin, myoglobin, ifosfamide, heavy metals, methotrexate, radiopaque contrast agents, streptozotocin Acute glomerulonephritis ANCA-associated: Crescentic glomerulonephritis, polyarteritis nodosa, Wegener's granulomatosis; Anti- GBM glomerulonephritis: Goodpasture's syndrome; Immune-complex: Lupus glomerulonephritis, postinfectious glomerulonephritis, cryoglobulinemic glomerulonephritis Acute tubulointerstitial Drug reaction (eg, β-lactams, NSAIDs, sulfonamides, nephritis ciprofloxacin, thiazide diuretics, furosemide, phenytoin, allopurinol, pyelonephritis, papillary necrosis Acute vascular Vasculitis, malignant hypertension, thrombotic nephropathy microangiopathies, scleroderma, atheroembolism Infiltrative diseases Lymphoma, sarcoidosis, leukemia Postrenal Tubular precipitation Uric acid (tumor lysis), sulfonamides, triamterene, acyclovir, indinavir, methotrexate, ethylene glycol ingestion, myeloma protein, myoglobin Ureteral obstruction Intrinsic: Calculi, clots, sloughed renal tissue, fungus ball, edema, malignancy, congenital defects; Extrinsic: Malignancy, retroperitoneal fibrosis, ureteral trauma during surgery or high impact injury Bladder obstruction Mechanical: Benign prostatic hyperplasia, prostate cancer, bladder cancer, urethral strictures, phimosis, paraphimosis, urethral valves, obstructed indwelling urinary catheter; Neurogenic: Anticholinergic drugs, upper or lower motor neuron lesion

In the case of ischemic ARF, the course of the disease may be divided into four phases. During an initiation phase, which lasts hours to days, reduced perfusion of the kidney is evolving into injury. Glomerular ultrafiltration reduces, the flow of filtrate is reduced due to debris within the tubules, and back leakage of filtrate through injured epithelium occurs. Renal injury can be mediated during this phase by reperfusion of the kidney. Initiation is followed by an extension phase which is characterized by continued ischemic injury and inflammation and may involve endothelial damage and vascular congestion. During the maintenance phase, lasting from 1 to 2 weeks, renal cell injury occurs, and glomerular filtration and urine output reaches a minimum. A recovery phase can follow in which the renal epithelium is repaired and GFR gradually recovers. Despite this, the survival rate of subjects with ARF may be as low as about 60%.

Acute kidney injury caused by radiocontrast agents (also called contrast media) and other nephrotoxins such as cyclosporine, antibiotics including aminoglycosides and anticancer drugs such as cisplatin manifests over a period of days to about a week. Contrast induced nephropathy (CIN, which is AKI caused by radiocontrast agents) is thought to be caused by intrarenal vasoconstriction (leading to ischemic injury) and from the generation of reactive oxygen species that are directly toxic to renal tubular epithelial cells. CIN classically presents as an acute (onset within 24-48 h) but reversible (peak 3-5 days, resolution within 1 week) rise in blood urea nitrogen and serum creatinine.

A commonly reported criteria for defining and detecting AKI is an abrupt (typically within about 2-7 days or within a period of hospitalization) elevation of serum creatinine. Although the use of serum creatinine elevation to define and detect AKI is well established, the magnitude of the serum creatinine elevation and the time over which it is measured to define AKI varies considerably among publications. Traditionally, relatively large increases in serum creatinine such as 100%, 200%, an increase of at least 100% to a value over 2 mg/dL and other definitions were used to define AKI. However, the recent trend has been towards using smaller serum creatinine rises to define AKI. The relationship between serum creatinine rise, AKI and the associated health risks are reviewed in Praught and Shlipak, Curr Opin Nephrol Hypertens 14:265-270, 2005 and Chertow et al, J Am Soc Nephrol 16: 3365-3370, 2005, which, with the references listed therein, are hereby incorporated by reference in their entirety. As described in these publications, acute worsening renal function (AKI) and increased risk of death and other detrimental outcomes are now known to be associated with very small increases in serum creatinine. These increases may be determined as a relative (percent) value or a nominal value. Relative increases in serum creatinine as small as 20% from the pre-injury value have been reported to indicate acutely worsening renal function (AKI) and increased health risk, but the more commonly reported value to define AKI and increased health risk is a relative increase of at least 25%. Nominal increases as small as 0.3 mg/dL, 0.2 mg/dL or even 0.1 mg/dL have been reported to indicate worsening renal function and increased risk of death. Various time periods for the serum creatinine to rise to these threshold values have been used to define AKI, for example, ranging from 2 days, 3 days, 7 days, or a variable period defined as the time the patient is in the hospital or intensive care unit. These studies indicate there is not a particular threshold serum creatinine rise (or time period for the rise) for worsening renal function or AKI, but rather a continuous increase in risk with increasing magnitude of serum creatinine rise.

One study (Lassnigg et all, J Am Soc Nephrol 15:1597-1605, 2004, hereby incorporated by reference in its entirety) investigated both increases and decreases in serum creatinine. Patients with a mild fall in serum creatinine of −0.1 to −0.3 mg/dL following heart surgery had the lowest mortality rate. Patients with a larger fall in serum creatinine (more than or equal to −0.4 mg/dL) or any increase in serum creatinine had a larger mortality rate. These findings caused the authors to conclude that even very subtle changes in renal function (as detected by small creatinine changes within 48 hours of surgery) seriously effect patient's outcomes. In an effort to reach consensus on a unified classification system for using serum creatinine to define AKI in clinical trials and in clinical practice, Bellomo et al., Crit Care. 8(4):R204-12, 2004, which is hereby incorporated by reference in its entirety, proposes the following classifications for stratifying AKI patients:

“Risk”: serum creatinine increased 1.5 fold from baseline OR urine production of <0.5 ml/kg body weight/hr for 6 hours; “Injury”: serum creatinine increased 2.0 fold from baseline OR urine production <0.5 ml/kg/hr for 12 h; “Failure”: serum creatinine increased 3.0 fold from baseline OR creatinine >355 μmol/1 (with a rise of >44) or urine output below 0.3 ml/kg/hr for 24 h or anuria for at least 12 hours;

And Included Two Clinical Outcomes:

“Loss”: persistent need for renal replacement therapy for more than four weeks. “ESRD”: end stage renal disease—the need for dialysis for more than 3 months. These criteria are called the RIFLE criteria, which provide a useful clinical tool to classify renal status. As discussed in Kellum, Crit. Care Med. 36: S141-45, 2008 and Ricci et al., Kidney Int. 73, 538-546, 2008, each hereby incorporated by reference in its entirety, the RIFLE criteria provide a uniform definition of AKI which has been validated in numerous studies.

More recently, Mehta et al., Crit. Care 11:R31 (doi:10.1186.cc5713), 2007, hereby incorporated by reference in its entirety, proposes the following similar classifications for stratifying AKI patients, which have been modified from RIFLE:

“Stage I”: increase in serum creatinine of more than or equal to 0.3 mg/dL (≥26.4 μmol/L) or increase to more than or equal to 150% (1.5-fold) from baseline OR urine output less than 0.5 mL/kg per hour for more than 6 hours; “Stage II”: increase in serum creatinine to more than 200% (>2-fold) from baseline OR urine output less than 0.5 mL/kg per hour for more than 12 hours; “Stage III”: increase in serum creatinine to more than 300% (>3-fold) from baseline OR serum creatinine ≥354 μmol/L accompanied by an acute increase of at least 44 μmol/L OR urine output less than 0.3 mL/kg per hour for 24 hours or anuria for 12 hours.

The CIN Consensus Working Panel (McCollough et al, Rev Cardiovasc Med. 2006; 7(4):177-197, hereby incorporated by reference in its entirety) uses a serum creatinine rise of 25% to define Contrast induced nephropathy (which is a type of AKI). Although various groups propose slightly different criteria for using serum creatinine to detect AKI, the consensus is that small changes in serum creatinine, such as 0.3 mg/dL or 25%, are sufficient to detect AKI (worsening renal function) and that the magnitude of the serum creatinine change is an indicator of the severity of the AKI and mortality risk.

Although serial measurement of serum creatinine over a period of days is an accepted method of detecting and diagnosing AKI and is considered one of the most important tools to evaluate AKI patients, serum creatinine is generally regarded to have several limitations in the diagnosis, assessment and monitoring of AKI patients. The time period for serum creatinine to rise to values (e.g., a 0.3 mg/dL or 25% rise) considered diagnostic for AKI can be 48 hours or longer depending on the definition used. Since cellular injury in AKI can occur over a period of hours, serum creatinine elevations detected at 48 hours or longer can be a late indicator of injury, and relying on serum creatinine can thus delay diagnosis of AKI. Furthermore, serum creatinine is not a good indicator of the exact kidney status and treatment needs during the most acute phases of AKI when kidney function is changing rapidly. Some patients with AKI will recover fully, some will need dialysis (either short term or long term) and some will have other detrimental outcomes including death, major adverse cardiac events and chronic kidney disease. Because serum creatinine is a marker of filtration rate, it does not differentiate between the causes of AKI (pre-renal, intrinsic renal, post-renal obstruction, atheroembolic, etc) or the category or location of injury in intrinsic renal disease (for example, tubular, glomerular or interstitial in origin). Urine output is similarly limited, Knowing these things can be of vital importance in managing and treating patients with AKI.

These limitations underscore the need for better methods to detect and assess AKI, particularly in the early and subclinical stages, but also in later stages when recovery and repair of the kidney can occur. Furthermore, there is a need to better identify patients who are at risk of having an AKI.

BRIEF SUMMARY OF THE INVENTION

It is an object of the invention to provide methods and compositions for evaluating renal function in a subject. As described herein, measurement of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24 (collectively referred to herein as “kidney injury markers, and individually as a “kidney injury marker”) can be used for diagnosis, prognosis, risk stratification, staging, monitoring, categorizing and determination of further diagnosis and treatment regimens in subjects suffering or at risk of suffering from an injury to renal function, reduced renal function, and/or acute renal failure (also called acute kidney injury).

These kidney injury markers may be used, individually or in panels comprising a plurality of kidney injury markers, for risk stratification (that is, to identify subjects at risk for a future injury to renal function, for future progression to reduced renal function, for future progression to ARF, for future improvement in renal function, etc.); for diagnosis of existing disease (that is, to identify subjects who have suffered an injury to renal function, who have progressed to reduced renal function, who have progressed to ARF, etc.); for monitoring for deterioration or improvement of renal function; and for predicting a future medical outcome, such as improved or worsening renal function, a decreased or increased mortality risk, a decreased or increased risk that a subject will require renal replacement therapy (i.e., hemodialysis, peritoneal dialysis, hemofiltration, and/or renal transplantation, a decreased or increased risk that a subject will recover from an injury to renal function, a decreased or increased risk that a subject will recover from ARF, a decreased or increased risk that a subject will progress to end stage renal disease, a decreased or increased risk that a subject will progress to chronic renal failure, a decreased or increased risk that a subject will suffer rejection of a transplanted kidney, etc.

In a first aspect, the present invention relates to methods for evaluating renal status in a subject. These methods comprise performing an assay method that is configured to detect one or more kidney injury markers of the present invention in a body fluid sample obtained from the subject. The assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are then correlated to the renal status of the subject. This correlation to renal status may include correlating the assay result(s) to one or more of risk stratification, diagnosis, prognosis, staging, classifying and monitoring of the subject as described herein. Thus, the present invention utilizes one or more kidney injury markers of the present invention for the evaluation of renal injury.

In certain embodiments, the methods for evaluating renal status described herein are methods for risk stratification of the subject; that is, assigning a likelihood of one or more future changes in renal status to the subject. In these embodiments, the assay result(s) is/are correlated to one or more such future changes. The following are preferred risk stratification embodiments.

In preferred risk stratification embodiments, these methods comprise determining a subject's risk for a future injury to renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a likelihood of such a future injury to renal function. For example, the measured concentration(s) may each be compared to a threshold value. For a “positive going” kidney injury marker, an increased likelihood of suffering a future injury to renal function is assigned to the subject when the measured concentration is above the threshold, relative to a likelihood assigned when the measured concentration is below the threshold. For a “negative going” kidney injury marker, an increased likelihood of suffering a future injury to renal function is assigned to the subject when the measured concentration is below the threshold, relative to a likelihood assigned when the measured concentration is above the threshold.

In other preferred risk stratification embodiments, these methods comprise determining a subject's risk for future reduced renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a likelihood of such reduced renal function. For example, the measured concentrations may each be compared to a threshold value. For a “positive going” kidney injury marker, an increased likelihood of suffering a future reduced renal function is assigned to the subject when the measured concentration is above the threshold, relative to a likelihood assigned when the measured concentration is below the threshold. For a “negative going” kidney injury marker, an increased likelihood of future reduced renal function is assigned to the subject when the measured concentration is below the threshold, relative to a likelihood assigned when the measured concentration is above the threshold.

In still other preferred risk stratification embodiments, these methods comprise determining a subject's likelihood for a future improvement in renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a likelihood of such a future improvement in renal function. For example, the measured concentration(s) may each be compared to a threshold value. For a “positive going” kidney injury marker, an increased likelihood of a future improvement in renal function is assigned to the subject when the measured concentration is below the threshold, relative to a likelihood assigned when the measured concentration is above the threshold. For a “negative going” kidney injury marker, an increased likelihood of a future improvement in renal function is assigned to the subject when the measured concentration is above the threshold, relative to a likelihood assigned when the measured concentration is below the threshold.

In yet other preferred risk stratification embodiments, these methods comprise determining a subject's risk for progression to ARF, and the result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a likelihood of such progression to ARF. For example, the measured concentration(s) may each be compared to a threshold value. For a “positive going” kidney injury marker, an increased likelihood of progression to ARF is assigned to the subject when the measured concentration is above the threshold, relative to a likelihood assigned when the measured concentration is below the threshold. For a “negative going” kidney injury marker, an increased likelihood of progression to ARF is assigned to the subject when the measured concentration is below the threshold, relative to a likelihood assigned when the measured concentration is above the threshold.

And in other preferred risk stratification embodiments, these methods comprise determining a subject's outcome risk, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a likelihood of the occurrence of a clinical outcome related to a renal injury suffered by the subject. For example, the measured concentration(s) may each be compared to a threshold value. For a “positive going” kidney injury marker, an increased likelihood of one or more of: acute kidney injury, progression to a worsening stage of AKI, mortality, a requirement for renal replacement therapy, a requirement for withdrawal of renal toxins, end stage renal disease, heart failure, stroke, myocardial infarction, progression to chronic kidney disease, etc., is assigned to the subject when the measured concentration is above the threshold, relative to a likelihood assigned when the measured concentration is below the threshold. For a “negative going” kidney injury marker, an increased likelihood of one or more of: acute kidney injury, progression to a worsening stage of AKI, mortality, a requirement for renal replacement therapy, a requirement for withdrawal of renal toxins, end stage renal disease, heart failure, stroke, myocardial infarction, progression to chronic kidney disease, etc., is assigned to the subject when the measured concentration is below the threshold, relative to a likelihood assigned when the measured concentration is above the threshold.

In such risk stratification embodiments, preferably the likelihood or risk assigned is that an event of interest is more or less likely to occur within 180 days of the time at which the body fluid sample is obtained from the subject. In particularly preferred embodiments, the likelihood or risk assigned relates to an event of interest occurring within a shorter time period such as 18 months, 120 days, 90 days, 60 days, 45 days, 30 days, 21 days, 14 days, 7 days, 5 days, 96 hours, 72 hours, 48 hours, 36 hours, 24 hours, 12 hours, or less. A risk at 0 hours of the time at which the body fluid sample is obtained from the subject is equivalent to diagnosis of a current condition.

In preferred risk stratification embodiments, the subject is selected for risk stratification based on the pre-existence in the subject of one or more known risk factors for prerenal, intrinsic renal, or postrenal ARF. For example, a subject undergoing or having undergone major vascular surgery, coronary artery bypass, or other cardiac surgery; a subject having pre-existing congestive heart failure, preeclampsia, eclampsia, diabetes mellitus, hypertension, coronary artery disease, proteinuria, renal insufficiency, glomerular filtration below the normal range, cirrhosis, serum creatinine above the normal range, or sepsis; or a subject exposed to NSAIDs, cyclosporines, tacrolimus, aminoglycosides, foscarnet, ethylene glycol, hemoglobin, myoglobin, ifosfamide, heavy metals, methotrexate, radiopaque contrast agents, or streptozotocin are all preferred subjects for monitoring risks according to the methods described herein. This list is not meant to be limiting. By “pre-existence” in this context is meant that the risk factor exists at the time the body fluid sample is obtained from the subject. In particularly preferred embodiments, a subject is chosen for risk stratification based on an existing diagnosis of injury to renal function, reduced renal function, or ARF.

In other embodiments, the methods for evaluating renal status described herein are methods for diagnosing a renal injury in the subject; that is, assessing whether or not a subject has suffered from an injury to renal function, reduced renal function, or ARF. In these embodiments, the assay result(s) is/are correlated to the occurrence or nonoccurrence of a change in renal status. The following are preferred diagnostic embodiments.

In preferred diagnostic embodiments, these methods comprise diagnosing the occurrence or nonoccurrence of an injury to renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of such an injury. For example, each of the measured concentration(s) may be compared to a threshold value. For a positive going marker, an increased likelihood of the occurrence of an injury to renal function is assigned to the subject when the measured concentration is above the threshold (relative to the likelihood assigned when the measured concentration is below the threshold); alternatively, when the measured concentration is below the threshold, an increased likelihood of the nonoccurrence of an injury to renal function may be assigned to the subject (relative to the likelihood assigned when the measured concentration is above the threshold). For a negative going marker, an increased likelihood of the occurrence of an injury to renal function is assigned to the subject when the measured concentration is below the threshold (relative to the likelihood assigned when the measured concentration is above the threshold); alternatively, when the measured concentration is above the threshold, an increased likelihood of the nonoccurrence of an injury to renal function may be assigned to the subject (relative to the likelihood assigned when the measured concentration is below the threshold).

In other preferred diagnostic embodiments, these methods comprise diagnosing the occurrence or nonoccurrence of reduced renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of an injury causing reduced renal function. For example, each of the measured concentration(s) may be compared to a threshold value. For a positive going marker, an increased likelihood of the occurrence of an injury causing reduced renal function is assigned to the subject when the measured concentration is above the threshold (relative to the likelihood assigned when the measured concentration is below the threshold); alternatively, when the measured concentration is below the threshold, an increased likelihood of the nonoccurrence of an injury causing reduced renal function may be assigned to the subject (relative to the likelihood assigned when the measured concentration is above the threshold). For a negative going marker, an increased likelihood of the occurrence of an injury causing reduced renal function is assigned to the subject when the measured concentration is below the threshold (relative to the likelihood assigned when the measured concentration is above the threshold); alternatively, when the measured concentration is above the threshold, an increased likelihood of the nonoccurrence of an injury causing reduced renal function may be assigned to the subject (relative to the likelihood assigned when the measured concentration is below the threshold).

In yet other preferred diagnostic embodiments, these methods comprise diagnosing the occurrence or nonoccurrence of ARF, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of an injury causing ARF. For example, each of the measured concentration(s) may be compared to a threshold value. For a positive going marker, an increased likelihood of the occurrence of ARF is assigned to the subject when the measured concentration is above the threshold (relative to the likelihood assigned when the measured concentration is below the threshold); alternatively, when the measured concentration is below the threshold, an increased likelihood of the nonoccurrence of ARF may be assigned to the subject (relative to the likelihood assigned when the measured concentration is above the threshold). For a negative going marker, an increased likelihood of the occurrence of ARF is assigned to the subject when the measured concentration is below the threshold (relative to the likelihood assigned when the measured concentration is above the threshold); alternatively, when the measured concentration is above the threshold, an increased likelihood of the nonoccurrence of ARF may be assigned to the subject (relative to the likelihood assigned when the measured concentration is below the threshold).

In still other preferred diagnostic embodiments, these methods comprise diagnosing a subject as being in need of renal replacement therapy, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a need for renal replacement therapy. For example, each of the measured concentration(s) may be compared to a threshold value. For a positive going marker, an increased likelihood of the occurrence of an injury creating a need for renal replacement therapy is assigned to the subject when the measured concentration is above the threshold (relative to the likelihood assigned when the measured concentration is below the threshold); alternatively, when the measured concentration is below the threshold, an increased likelihood of the nonoccurrence of an injury creating a need for renal replacement therapy may be assigned to the subject (relative to the likelihood assigned when the measured concentration is above the threshold). For a negative going marker, an increased likelihood of the occurrence of an injury creating a need for renal replacement therapy is assigned to the subject when the measured concentration is below the threshold (relative to the likelihood assigned when the measured concentration is above the threshold); alternatively, when the measured concentration is above the threshold, an increased likelihood of the nonoccurrence of an injury creating a need for renal replacement therapy may be assigned to the subject (relative to the likelihood assigned when the measured concentration is below the threshold).

In still other preferred diagnostic embodiments, these methods comprise diagnosing a subject as being in need of renal transplantation, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a need for renal transplantation. For example, each of the measured concentration(s) may be compared to a threshold value. For a positive going marker, an increased likelihood of the occurrence of an injury creating a need for renal transplantation is assigned to the subject when the measured concentration is above the threshold (relative to the likelihood assigned when the measured concentration is below the threshold); alternatively, when the measured concentration is below the threshold, an increased likelihood of the nonoccurrence of an injury creating a need for renal transplantation may be assigned to the subject (relative to the likelihood assigned when the measured concentration is above the threshold). For a negative going marker, an increased likelihood of the occurrence of an injury creating a need for renal transplantation is assigned to the subject when the measured concentration is below the threshold (relative to the likelihood assigned when the measured concentration is above the threshold); alternatively, when the measured concentration is above the threshold, an increased likelihood of the nonoccurrence of an injury creating a need for renal transplantation may be assigned to the subject (relative to the likelihood assigned when the measured concentration is below the threshold).

In still other embodiments, the methods for evaluating renal status described herein are methods for monitoring a renal injury in the subject; that is, assessing whether or not renal function is improving or worsening in a subject who has suffered from an injury to renal function, reduced renal function, or ARF. In these embodiments, the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of a change in renal status. The following are preferred monitoring embodiments.

In preferred monitoring embodiments, these methods comprise monitoring renal status in a subject suffering from an injury to renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of a change in renal status in the subject. For example, the measured concentration(s) may be compared to a threshold value. For a positive going marker, when the measured concentration is above the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is below the threshold, an improvement of renal function may be assigned to the subject. For a negative going marker, when the measured concentration is below the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is above the threshold, an improvement of renal function may be assigned to the subject.

In other preferred monitoring embodiments, these methods comprise monitoring renal status in a subject suffering from reduced renal function, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of a change in renal status in the subject. For example, the measured concentration(s) may be compared to a threshold value. For a positive going marker, when the measured concentration is above the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is below the threshold, an improvement of renal function may be assigned to the subject. For a negative going marker, when the measured concentration is below the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is above the threshold, an improvement of renal function may be assigned to the subject.

In yet other preferred monitoring embodiments, these methods comprise monitoring renal status in a subject suffering from acute renal failure, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of a change in renal status in the subject. For example, the measured concentration(s) may be compared to a threshold value. For a positive going marker, when the measured concentration is above the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is below the threshold, an improvement of renal function may be assigned to the subject. For a negative going marker, when the measured concentration is below the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is above the threshold, an improvement of renal function may be assigned to the subject.

In other additional preferred monitoring embodiments, these methods comprise monitoring renal status in a subject at risk of an injury to renal function due to the pre-existence of one or more known risk factors for prerenal, intrinsic renal, or postrenal ARF, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the occurrence or nonoccurrence of a change in renal status in the subject. For example, the measured concentration(s) may be compared to a threshold value. For a positive going marker, when the measured concentration is above the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is below the threshold, an improvement of renal function may be assigned to the subject. For a negative going marker, when the measured concentration is below the threshold, a worsening of renal function may be assigned to the subject; alternatively, when the measured concentration is above the threshold, an improvement of renal function may be assigned to the subject.

In yet other preferred monitoring embodiments, these methods comprise monitoring renal status in a subject having, or at risk of, an injury to renal function for future persistence of acute kidney injury. “Future persistence” as used herein refers to an existing acute renal injury that will continue for a period selected from the group consisting of 21 days, 14 days, 7 days, 5 days, 96 hours, 72 hours, 48 hours, 36 hours, 24 hours, and 12 hours. In certain embodiments the subject has an acute kidney injury at the time the sample is obtained. This is not meant to imply that the subject must have an acute kidney injury at the time the sample is obtained, but rather that the subject, upon onset of an acute kidney injury, suffers from an acute kidney injury that will persist. In various embodiments, the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the future persistence of the acute kidney injury in the subject. For example, the measured concentration(s) may be compared to a threshold value. For a positive going marker, when the measured concentration is above the threshold, a future persistence of acute kidney injury may be assigned to the subject; alternatively, when the measured concentration is below the threshold, a future improvement of renal function may be assigned to the subject. For a negative going marker, when the measured concentration is below the threshold, a future persistence of acute kidney injury may be assigned to the subject; alternatively, when the measured concentration is above the threshold, a future improvement of renal function may be assigned to the subject.

In still other embodiments, the methods for evaluating renal status described herein are methods for classifying a renal injury in the subject; that is, determining whether a renal injury in a subject is prerenal, intrinsic renal, or postrenal; and/or further subdividing these classes into subclasses such as acute tubular injury, acute glomerulonephritis acute tubulointerstitial nephritis, acute vascular nephropathy, or infiltrative disease; and/or assigning a likelihood that a subject will progress to a particular RIFLE stage. In these embodiments, the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to a particular class and/or subclass. The following are preferred classification embodiments.

In preferred classification embodiments, these methods comprise determining whether a renal injury in a subject is prerenal, intrinsic renal, or postrenal; and/or further subdividing these classes into subclasses such as acute tubular injury, acute glomerulonephritis acute tubulointerstitial nephritis, acute vascular nephropathy, or infiltrative disease; and/or assigning a likelihood that a subject will progress to a particular RIFLE stage, and the assay result(s), for example a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24, is/are correlated to the injury classification for the subject. For example, the measured concentration may be compared to a threshold value, and when the measured concentration is above the threshold, a particular classification is assigned; alternatively, when the measured concentration is below the threshold, a different classification may be assigned to the subject.

A variety of methods may be used by the skilled artisan to arrive at a desired threshold value for use in these methods. For example, the threshold value may be determined from a population of normal subjects by selecting a concentration representing the 75^(th), 85^(th), 90^(th), 95^(th), or 99^(th) percentile of a kidney injury marker measured in such normal subjects. Alternatively, the threshold value may be determined from a “diseased” population of subjects, e.g., those suffering from an injury or having a predisposition for an injury (e.g., progression to ARF or some other clinical outcome such as death, dialysis, renal transplantation, etc.), by selecting a concentration representing the 75^(th), 85^(th), 90^(th), 95^(th), or 99^(th) percentile of a kidney injury marker measured in such subjects. In another alternative, the threshold value may be determined from a prior measurement of a kidney injury marker in the same subject; that is, a temporal change in the level of a kidney injury marker in the subject may be used to assign risk to the subject.

The foregoing discussion is not meant to imply, however, that the kidney injury markers of the present invention must be compared to corresponding individual thresholds. Methods for combining assay results can comprise the use of multivariate logistical regression, loglinear modeling, neural network analysis, n-of-m analysis, decision tree analysis, calculating ratios of markers, etc. This list is not meant to be limiting. In these methods, a composite result which is determined by combining individual markers may be treated as if it is itself a marker; that is, a threshold may be determined for the composite result as described herein for individual markers, and the composite result for an individual patient compared to this threshold.

The ability of a particular test to distinguish two populations can be established using ROC analysis. For example, ROC curves established from a “first” subpopulation which is predisposed to one or more future changes in renal status, and a “second” subpopulation which is not so predisposed can be used to calculate a ROC curve, and the area under the curve provides a measure of the quality of the test. Preferably, the tests described herein provide a ROC curve area greater than 0.5, preferably at least 0.6, more preferably 0.7, still more preferably at least 0.8, even more preferably at least 0.9, and most preferably at least 0.95.

In certain aspects, the measured concentration of one or more kidney injury markers, or a composite of such markers, may be treated as continuous variables. For example, any particular concentration can be converted into a corresponding probability of a future reduction in renal function for the subject, the occurrence of an injury, a classification, etc. In yet another alternative, a threshold that can provide an acceptable level of specificity and sensitivity in separating a population of subjects into “bins” such as a “first” subpopulation (e.g., which is predisposed to one or more future changes in renal status, the occurrence of an injury, a classification, etc.) and a “second” subpopulation which is not so predisposed. A threshold value is selected to separate this first and second population by one or more of the following measures of test accuracy:

an odds ratio greater than 1, preferably at least about 2 or more or about 0.5 or less, more preferably at least about 3 or more or about 0.33 or less, still more preferably at least about 4 or more or about 0.25 or less, even more preferably at least about 5 or more or about 0.2 or less, and most preferably at least about 10 or more or about 0.1 or less; a specificity of greater than 0.5, preferably at least about 0.6, more preferably at least about 0.7, still more preferably at least about 0.8, even more preferably at least about 0.9 and most preferably at least about 0.95, with a corresponding sensitivity greater than 0.2, preferably greater than about 0.3, more preferably greater than about 0.4, still more preferably at least about 0.5, even more preferably about 0.6, yet more preferably greater than about 0.7, still more preferably greater than about 0.8, more preferably greater than about 0.9, and most preferably greater than about 0.95; a sensitivity of greater than 0.5, preferably at least about 0.6, more preferably at least about 0.7, still more preferably at least about 0.8, even more preferably at least about 0.9 and most preferably at least about 0.95, with a corresponding specificity greater than 0.2, preferably greater than about 0.3, more preferably greater than about 0.4, still more preferably at least about 0.5, even more preferably about 0.6, yet more preferably greater than about 0.7, still more preferably greater than about 0.8, more preferably greater than about 0.9, and most preferably greater than about 0.95; at least about 75% sensitivity, combined with at least about 75% specificity; a positive likelihood ratio (calculated as sensitivity/(1-specificity)) of greater than 1, at least about 2, more preferably at least about 3, still more preferably at least about 5, and most preferably at least about 10; or a negative likelihood ratio (calculated as (1-sensitivity)/specificity) of less than 1, less than or equal to about 0.5, more preferably less than or equal to about 0.3, and most preferably less than or equal to about 0.1. The term “about” in the context of any of the above measurements refers to +/−5% of a given measurement.

Multiple thresholds may also be used to assess renal status in a subject. For example, a “first” subpopulation which is predisposed to one or more future changes in renal status, the occurrence of an injury, a classification, etc., and a “second” subpopulation which is not so predisposed can be combined into a single group. This group is then subdivided into three or more equal parts (known as tertiles, quartiles, quintiles, etc., depending on the number of subdivisions). An odds ratio is assigned to subjects based on which subdivision they fall into. If one considers a tertile, the lowest or highest tertile can be used as a reference for comparison of the other subdivisions. This reference subdivision is assigned an odds ratio of 1. The second tertile is assigned an odds ratio that is relative to that first tertile. That is, someone in the second tertile might be 3 times more likely to suffer one or more future changes in renal status in comparison to someone in the first tertile. The third tertile is also assigned an odds ratio that is relative to that first tertile.

In certain embodiments, the assay method is an immunoassay. Antibodies for use in such assays will specifically bind a full length kidney injury marker of interest, and may also bind one or more polypeptides that are “related” thereto, as that term is defined hereinafter. Numerous immunoassay formats are known to those of skill in the art. Preferred body fluid samples are selected from the group consisting of urine, blood, serum, saliva, tears, and plasma.

The foregoing method steps should not be interpreted to mean that the kidney injury marker assay result(s) is/are used in isolation in the methods described herein. Rather, additional variables or other clinical indicia may be included in the methods described herein. For example, a risk stratification, diagnostic, classification, monitoring, etc. method may combine the assay result(s) with one or more variables measured for the subject selected from the group consisting of demographic information (e.g., weight, sex, age, race), medical history (e.g., family history, type of surgery, pre-existing disease such as aneurism, congestive heart failure, preeclampsia, eclampsia, diabetes mellitus, hypertension, coronary artery disease, proteinuria, renal insufficiency, or sepsis, type of toxin exposure such as NSAIDs, cyclosporines, tacrolimus, aminoglycosides, foscarnet, ethylene glycol, hemoglobin, myoglobin, ifosfamide, heavy metals, methotrexate, radiopaque contrast agents, or streptozotocin), clinical variables (e.g., blood pressure, temperature, respiration rate), risk scores (APACHE score, PREDICT score, TIMI Risk Score for UA/NSTEMI, Framingham Risk Score), a glomerular filtration rate, an estimated glomerular filtration rate, a urine production rate, a serum or plasma creatinine concentration, a urine creatinine concentration, a fractional excretion of sodium, a urine sodium concentration, a urine creatinine to serum or plasma creatinine ratio, a urine specific gravity, a urine osmolality, a urine urea nitrogen to plasma urea nitrogen ratio, a plasma BUN to creatinine ratio, a renal failure index calculated as urine sodium/(urine creatinine/plasma creatinine), a serum or plasma neutrophil gelatinase (NGAL) concentration, a urine NGAL concentration, a serum or plasma cystatin C concentration, a serum or plasma cardiac troponin concentration, a serum or plasma BNP concentration, a serum or plasma NTproBNP concentration, and a serum or plasma proBNP concentration. Other measures of renal function which may be combined with one or more kidney injury marker assay result(s) are described hereinafter and in Harrison's Principles of Internal Medicine, 17^(th) Ed., McGraw Hill, New York, pages 1741-1830, and Current Medical Diagnosis & Treatment 2008, 47^(th) Ed, McGraw Hill, New York, pages 785-815, each of which are hereby incorporated by reference in their entirety.

When more than one marker is measured, the individual markers may be measured in samples obtained at the same time, or may be determined from samples obtained at different (e.g., an earlier or later) times. The individual markers may also be measured on the same or different body fluid samples. For example, one kidney injury marker may be measured in a serum or plasma sample and another kidney injury marker may be measured in a urine sample. In addition, assignment of a likelihood may combine an individual kidney injury marker assay result with temporal changes in one or more additional variables.

In various related aspects, the present invention also relates to devices and kits for performing the methods described herein. Suitable kits comprise reagents sufficient for performing an assay for at least one of the described kidney injury markers, together with instructions for performing the described threshold comparisons.

In certain embodiments, reagents for performing such assays are provided in an assay device, and such assay devices may be included in such a kit. Preferred reagents can comprise one or more solid phase antibodies, the solid phase antibody comprising antibody that detects the intended biomarker target(s) bound to a solid support. In the case of sandwich immunoassays, such reagents can also include one or more detectably labeled antibodies, the detectably labeled antibody comprising antibody that detects the intended biomarker target(s) bound to a detectable label. Additional optional elements that may be provided as part of an assay device are described hereinafter.

Detectable labels may include molecules that are themselves detectable (e.g., fluorescent moieties, electrochemical labels, ecl (electrochemical luminescence) labels, metal chelates, colloidal metal particles, etc.) as well as molecules that may be indirectly detected by production of a detectable reaction product (e.g., enzymes such as horseradish peroxidase, alkaline phosphatase, etc.) or through the use of a specific binding molecule which itself may be detectable (e.g., a labeled antibody that binds to the second antibody, biotin, digoxigenin, maltose, oligohistidine, 2,4-dinitrobenzene, phenylarsenate, ssDNA, dsDNA, etc.).

Generation of a signal from the signal development element can be performed using various optical, acoustical, and electrochemical methods well known in the art. Examples of detection modes include fluorescence, radiochemical detection, reflectance, absorbance, amperometry, conductance, impedance, interferometry, ellipsometry, etc. In certain of these methods, the solid phase antibody is coupled to a transducer (e.g., a diffraction grating, electrochemical sensor, etc) for generation of a signal, while in others, a signal is generated by a transducer that is spatially separate from the solid phase antibody (e.g., a fluorometer that employs an excitation light source and an optical detector). This list is not meant to be limiting. Antibody-based biosensors may also be employed to determine the presence or amount of analytes that optionally eliminate the need for a labeled molecule.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to methods and compositions for diagnosis, differential diagnosis, risk stratification, monitoring, classifying and determination of treatment regimens in subjects suffering or at risk of suffering from injury to renal function, reduced renal function and/or acute renal failure through measurement of one or more kidney injury markers. In various embodiments, a measured concentration of one or more markers selected from the group consisting of Metalloproteinase inhibitor 1, Metalloproteinase inhibitor 2, Metalloproteinase inhibitor 4, C—C motif chemokine 15, C—C motif chemokine 18, C—C motif chemokine 23, and/or, C—C motif chemokine 24 or one or more markers related thereto, and optionally one or more additional kidney injury markers known in the art, are correlated to the renal status of the subject.

For purposes of this document, the following definitions apply:

As used herein, an “injury to renal function” is an abrupt (within 14 days, preferably within 7 days, more preferably within 72 hours, and still more preferably within 48 hours) measurable reduction in a measure of renal function. Such an injury may be identified, for example, by a decrease in glomerular filtration rate or estimated GFR, a reduction in urine output, an increase in serum creatinine, an increase in serum cystatin C, a requirement for renal replacement therapy, etc. “Improvement in Renal Function” is an abrupt (within 14 days, preferably within 7 days, more preferably within 72 hours, and still more preferably within 48 hours) measurable increase in a measure of renal function. Preferred methods for measuring and/or estimating GFR are described hereinafter. As used herein, “reduced renal function” is an abrupt (within 14 days, preferably within 7 days, more preferably within 72 hours, and still more preferably within 48 hours) reduction in kidney function identified by an absolute increase in serum creatinine of greater than or equal to 0.1 mg/dL (≥8.8 μmol/L), a percentage increase in serum creatinine of greater than or equal to 20% (1.2-fold from baseline), or a reduction in urine output (documented oliguria of less than 0.5 ml/kg per hour). As used herein, “acute renal failure” or “ARF” is an abrupt (within 14 days, preferably within 7 days, more preferably within 72 hours, and still more preferably within 48 hours) reduction in kidney function identified by an absolute increase in serum creatinine of greater than or equal to 0.3 mg/dl (≥26.4 μmol/l), a percentage increase in serum creatinine of greater than or equal to 50% (1.5-fold from baseline), or a reduction in urine output (documented oliguria of less than 0.5 ml/kg per hour for at least 6 hours). This term is synonymous with “acute kidney injury” or “AKI.”

In this regard, the skilled artisan will understand that the signals obtained from an immunoassay are a direct result of complexes formed between one or more antibodies and the target biomolecule (i.e., the analyte) and polypeptides containing the necessary epitope(s) to which the antibodies bind. While such assays may detect the full length biomarker and the assay result be expressed as a concentration of a biomarker of interest, the signal from the assay is actually a result of all such “immunoreactive” polypeptides present in the sample. Expression of biomarkers may also be determined by means other than immunoassays, including protein measurements (such as dot blots, western blots, chromatographic methods, mass spectrometry, etc.) and nucleic acid measurements (mRNA quantitation). This list is not meant to be limiting.

As used herein, the term “Metalloproteinase inhibitor 1” refers to one or more polypeptides present in a biological sample that are derived from the Metalloproteinase inhibitor 1 precursor (Swiss-Prot P01033 (SEQ ID NO: 1)).

        10         20         30         40 MAPFEPLASG ILLLLWLIAP SRACTCVPPH PQTAFCNSDL         50         60         70         80 VIRAKFVGTP EVNQTTLYQR YEIKMTKMYK GFQALGDAAD         90        100        110        120 IRFVYTPAME SVCGYFHRSH NRSEEFLIAG KLQDGLLHIT        130        140        150        160 TCSFVAPWNS LSLAQRRGFT KTYTVGCEEC TVFPCLSIPC        170        180        190        200 KLQSGTHCLW TDQLLQGSEK GFQSRHLACL PREPGLCTWQ SLRSQIA

The following domains have been identified in Metalloproteinase inhibitor 1:

Residues Length Domain ID  1-23  23 signal sequence 24-207 184 Metalloproteinase inhibitor 1

As used herein, the term “Metalloproteinase inhibitor 2” refers to one or more polypeptides present in a biological sample that are derived from the Metalloproteinase inhibitor 2 precursor (human precursor: Swiss-Prot P16035 (SEQ ID NO: 2)).

        10         20         30         40 MGAAARTLRL ALGLLLLATL LRPADACSCS PVHPQQAFCN         50         60         70         80 ADVVIRAKAV SEKEVDSGND IYGNPIKRIQ YEIKQIKMFK         90        100        110        120 GPEKDIEFIY TAPSSAVCGV SLDVGGKKEY LIAGKAEGDG        130        140        150        160 KMHITLCDFI VPWDTLSTTQ KKSLNHRYQM GCECKITRCP        170        180        190        200 MIPCYISSPD ECLWMDWVTE KNINGHQAKF FACIKRSDGS        210        220 CAWYRGAAPP KQEFLDIEDP

The following domains have been identified in Metalloproteinase inhibitor 2:

Residues Length Domain ID  1-26  26 Signal peptide 27-220 194 Metalloproteinase inhibitor 2

As used herein, the term “Metalloproteinase inhibitor 4” refers to one or polypeptides present in a biological sample that are derived from the Metalloproteinase inhibitor 4 precursor (human precursor: Swiss-Prot Q99727 (SEQ ID NO: 3)).

        10         20         30         40 MPGSPRPAPS WVLLLRLLAL LRPPGLGEAC SCAPAHPQQH         50         60         70         80 ICHSALVIRA KISSEKVVPA SADPADTEKM LRYEIKQIKM         90        100        110        120 FKGFEKVKDV QYIYTPFDSS LCGVKLEANS QKQYLLTGQV        130        140        150        160 LSDGKVFIHL CNYIEPWEDL SLVQRESLNH HYHLNCGCQI        170        180        190        200 TTCYTVPCTI SAPNECLWTD WLLERKLYGY QAQHYVCMKH        210        220 VDGTCSWYRG HLPLRKEFVD IVQP

The following domains have been identified in Metalloproteinase inhibitor 4:

Residues Length Domain ID  1-29  29 Signal sequence 30-224 195 Metalloproteinase inhibitor 4

As used herein, the term “C—C motif chemokine 15” refers to one or more polypeptides present in a biological sample that are derived from the C—C motif chemokine 15 precursor (human precursor: Swiss-Prot Q16663 (SEQ ID NO: 4)).

        10         20         30         40 MKVSVAALSC LMLVAVLGSQ AQFINDAETE LMMSKLPLEN         50         60         70         80 PVVLNSFHFA ADCCTSYISQ SIPCSLMKSY FETSSECSKP         90        100        110 GVIFLTKKGR QVCAKPSGPG VQDCMKKLKP YSI

The following domains have been identified in C—C motif chemokine 15:

Residues Length Domain ID  1-21 21 Signal peptide 22-113 92 C—C motif chemokine 15 43-113 71 C—C motif chemokine 15 (22-92) 46-113 68 C—C motif chemokine 15 (25-92) 50-113 64 C—C motif chemokine 15 (29-92)

As used herein, the term “C—C motif chemokine 18” refers to one or more polypeptides present in a biological sample that are derived from the C—C motif chemokine 18 precursor (human precursor: Swiss-Prot P55774 (SEQ ID NO: 5)).

        10         20         30         40 MKGLAAALLV LVCTMALCSC AQVGTNKELC CLVYTSWQIP         50         60         70         80 QKFIVDYSET SPQCPKPGVI LLTKRGRQIC ADPNKKWVQK  YISDLKLNA

The following domains have been identified in C—C motif chemokine 18:

Residues Length Domain ID  1-20 20 Signal peptide 21-89 69 C—C motif chemokine 18 21-88 68 CCL 18 (1-68) 23-89 67 CCL 18 (3-69) 24-89 66 CCL 18 (4-69)

As used herein, the term “C—C motif chemokine 23” refers to one or more polypeptides present in a biological sample that are derived from the C—C motif chemokine 23 precursor (Swiss-Prot P55773 (SEQ ID NO: 6))

        10         20         30         40 MKVSVAALSC LMLVTALGSQ ARVTKDAETE FMMSKLPLEN         50         60         70         80 PVLLDRFHAT SADCCISYTP RSIPCSLLES YFETNSECSK         90        100        110        120 PGVIFLTKKG RRFCANPSDK QVQVCVRMLK LDTRIKTRKN

The following domains have been identified in C—C motif chemokine 23:

Residues Length Domain ID  1-21 21 signal peptide 22-120 99 C—C motif chemokine 23 40-120 81 C—C motif chemokine 23 (19-99) 43-120 78 C—C motif chemokine 23 (22-99) 48-120 73 C—C motif chemokine 23 (27-99) 51-120 70 C—C motif chemokine 23 (30-99)

As used herein, the term “C—C motif chemokine 24” refers to one or more polypeptides present in a biological sample that are derived from the C—C motif chemokine 24 precursor (Swiss-Prot 000175 (SEQ ID NO: 7))

        10         20        30         40 MAGLMTIVTS LLFLGVCAHH IIPTGSWIP SPCCMFFVSK        50         60         70         80 RIPENRWSY QLSSRSTCLK AGVIFTTKKG QQFCGDPKQE         90        100        110 WVQRYMKNLD AKQKKASPRA RAVAVKGPVQ RYPGNQTTC

The following domains have been identified in C—C motif chemokine 24:

Residues Length Domain ID  1-26 26 signal peptide 27-119 93 C—C motif chemokine 24

As used herein, the term “relating a signal to the presence or amount” of an analyte reflects this understanding. Assay signals are typically related to the presence or amount of an analyte through the use of a standard curve calculated using known concentrations of the analyte of interest. As the term is used herein, an assay is “configured to detect” an analyte if an assay can generate a detectable signal indicative of the presence or amount of a physiologically relevant concentration of the analyte. Because an antibody epitope is on the order of 8 amino acids, an immunoassay configured to detect a marker of interest will also detect polypeptides related to the marker sequence, so long as those polypeptides contain the epitope(s) necessary to bind to the antibody or antibodies used in the assay.

The term “related marker” as used herein with regard to a biomarker such as one of the kidney injury markers described herein refers to one or more fragments, variants, etc., of a particular marker or its biosynthetic parent that may be detected as a surrogate for the marker itself or as independent biomarkers. The term also refers to one or more polypeptides present in a biological sample that are derived from the biomarker precursor complexed to additional species, such as binding proteins, receptors, heparin, lipids, sugars, etc.

The term “positive going” marker as that term is used herein refer to a marker that is determined to be elevated in subjects suffering from a disease or condition, relative to subjects not suffering from that disease or condition. The term “negative going” marker as that term is used herein refer to a marker that is determined to be reduced in subjects suffering from a disease or condition, relative to subjects not suffering from that disease or condition.

The term “subject” as used herein refers to a human or non-human organism. Thus, the methods and compositions described herein are applicable to both human and veterinary disease. Further, while a subject is preferably a living organism, the invention described herein may be used in post-mortem analysis as well. Preferred subjects are humans, and most preferably “patients,” which as used herein refers to living humans that are receiving medical care for a disease or condition. This includes persons with no defined illness who are being investigated for signs of pathology.

Preferably, an analyte is measured in a sample. Such a sample may be obtained from a subject, or may be obtained from biological materials intended to be provided to the subject. For example, a sample may be obtained from a kidney being evaluated for possible transplantation into a subject, and an analyte measurement used to evaluate the kidney for preexisting damage. Preferred samples are body fluid samples.

The term “body fluid sample” as used herein refers to a sample of bodily fluid obtained for the purpose of diagnosis, prognosis, classification or evaluation of a subject of interest, such as a patient or transplant donor. In certain embodiments, such a sample may be obtained for the purpose of determining the outcome of an ongoing condition or the effect of a treatment regimen on a condition. Preferred body fluid samples include blood, serum, plasma, cerebrospinal fluid, urine, saliva, sputum, and pleural effusions. In addition, one of skill in the art would realize that certain body fluid samples would be more readily analyzed following a fractionation or purification procedure, for example, separation of whole blood into serum or plasma components.

The term “diagnosis” as used herein refers to methods by which the skilled artisan can estimate and/or determine the probability (“a likelihood”) of whether or not a patient is suffering from a given disease or condition. In the case of the present invention, “diagnosis” includes using the results of an assay, most preferably an immunoassay, for a kidney injury marker of the present invention, optionally together with other clinical characteristics, to arrive at a diagnosis (that is, the occurrence or nonoccurrence) of an acute renal injury or ARF for the subject from which a sample was obtained and assayed. That such a diagnosis is “determined” is not meant to imply that the diagnosis is 100% accurate. Many biomarkers are indicative of multiple conditions. The skilled clinician does not use biomarker results in an informational vacuum, but rather test results are used together with other clinical indicia to arrive at a diagnosis. Thus, a measured biomarker level on one side of a predetermined diagnostic threshold indicates a greater likelihood of the occurrence of disease in the subject relative to a measured level on the other side of the predetermined diagnostic threshold.

Similarly, a prognostic risk signals a probability (“a likelihood”) that a given course or outcome will occur. A level or a change in level of a prognostic indicator, which in turn is associated with an increased probability of morbidity (e.g., worsening renal function, future ARF, or death) is referred to as being “indicative of an increased likelihood” of an adverse outcome in a patient.

Marker Assays

In general, immunoassays involve contacting a sample containing or suspected of containing a biomarker of interest with at least one antibody that specifically binds to the biomarker. A signal is then generated indicative of the presence or amount of complexes formed by the binding of polypeptides in the sample to the antibody. The signal is then related to the presence or amount of the biomarker in the sample. Numerous methods and devices are well known to the skilled artisan for the detection and analysis of biomarkers. See, e.g., U.S. Pat. Nos. 6,143,576; 6,113,855; 6,019,944; 5,985,579; 5,947,124; 5,939,272; 5,922,615; 5,885,527; 5,851,776; 5,824,799; 5,679,526; 5,525,524; and 5,480,792, and The Immunoassay Handbook, David Wild, ed. Stockton Press, New York, 1994, each of which is hereby incorporated by reference in its entirety, including all tables, FIGURES and claims.

The assay devices and methods known in the art can utilize labeled molecules in various sandwich, competitive, or non-competitive assay formats, to generate a signal that is related to the presence or amount of the biomarker of interest. Suitable assay formats also include chromatographic, mass spectrographic, and protein “blotting” methods. Additionally, certain methods and devices, such as biosensors and optical immunoassays, may be employed to determine the presence or amount of analytes without the need for a labeled molecule. See, e.g., U.S. Pat. Nos. 5,631,171; and 5,955,377, each of which is hereby incorporated by reference in its entirety, including all tables, FIGURES and claims. One skilled in the art also recognizes that robotic instrumentation including but not limited to Beckman ACCESS®, Abbott AXSYM®, Roche ELECSYS®, Dade Behring STRATUS® systems are among the immunoassay analyzers that are capable of performing immunoassays. But any suitable immunoassay may be utilized, for example, enzyme-linked immunoassays (ELISA), radioimmunoassays (RIAs), competitive binding assays, and the like.

Antibodies or other polypeptides may be immobilized onto a variety of solid supports for use in assays. Solid phases that may be used to immobilize specific binding members include those developed and/or used as solid phases in solid phase binding assays. Examples of suitable solid phases include membrane filters, cellulose-based papers, beads (including polymeric, latex and paramagnetic particles), glass, silicon wafers, microparticles, nanoparticles, TentaGels, AgroGels, PEGA gels, SPOCC gels, and multiple-well plates. An assay strip could be prepared by coating the antibody or a plurality of antibodies in an array on solid support. This strip could then be dipped into the test sample and then processed quickly through washes and detection steps to generate a measurable signal, such as a colored spot. Antibodies or other polypeptides may be bound to specific zones of assay devices either by conjugating directly to an assay device surface, or by indirect binding. In an example of the later case, antibodies or other polypeptides may be immobilized on particles or other solid supports, and that solid support immobilized to the device surface.

Biological assays require methods for detection, and one of the most common methods for quantitation of results is to conjugate a detectable label to a protein or nucleic acid that has affinity for one of the components in the biological system being studied. Detectable labels may include molecules that are themselves detectable (e.g., fluorescent moieties, electrochemical labels, metal chelates, etc.) as well as molecules that may be indirectly detected by production of a detectable reaction product (e.g., enzymes such as horseradish peroxidase, alkaline phosphatase, etc.) or by a specific binding molecule which itself may be detectable (e.g., biotin, digoxigenin, maltose, oligohistidine, 2,4-dinitrobenzene, phenylarsenate, ssDNA, dsDNA, etc.).

Preparation of solid phases and detectable label conjugates often comprise the use of chemical cross-linkers. Cross-linking reagents contain at least two reactive groups, and are divided generally into homofunctional cross-linkers (containing identical reactive groups) and heterofunctional cross-linkers (containing non-identical reactive groups). Homobifunctional cross-linkers that couple through amines, sulfhydryls or react non-specifically are available from many commercial sources. Maleimides, alkyl and aryl halides, alpha-haloacyls and pyridyl disulfides are thiol reactive groups. Maleimides, alkyl and aryl halides, and alpha-haloacyls react with sulfhydryls to form thiol ether bonds, while pyridyl disulfides react with sulfhydryls to produce mixed disulfides. The pyridyl disulfide product is cleavable. Imidoesters are also very useful for protein-protein cross-links. A variety of heterobifunctional cross-linkers, each combining different attributes for successful conjugation, are commercially available.

In certain aspects, the present invention provides kits for the analysis of the described kidney injury markers. The kit comprises reagents for the analysis of at least one test sample which comprise at least one antibody that a kidney injury marker. The kit can also include devices and instructions for performing one or more of the diagnostic and/or prognostic correlations described herein. Preferred kits will comprise an antibody pair for performing a sandwich assay, or a labeled species for performing a competitive assay, for the analyte. Preferably, an antibody pair comprises a first antibody conjugated to a solid phase and a second antibody conjugated to a detectable label, wherein each of the first and second antibodies that bind a kidney injury marker. Most preferably each of the antibodies are monoclonal antibodies. The instructions for use of the kit and performing the correlations can be in the form of labeling, which refers to any written or recorded material that is attached to, or otherwise accompanies a kit at any time during its manufacture, transport, sale or use. For example, the term labeling encompasses advertising leaflets and brochures, packaging materials, instructions, audio or video cassettes, computer discs, as well as writing imprinted directly on kits.

Antibodies

The term “antibody” as used herein refers to a peptide or polypeptide derived from, modeled after or substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof, capable of specifically binding an antigen or epitope. See, e.g. Fundamental Immunology, 3rd Edition, W. E. Paul, ed., Raven Press, N.Y. (1993); Wilson (1994; J. Immunol. Methods 175:267-273; Yarmush (1992) J. Biochem. Biophys. Methods 25:85-97. The term antibody includes antigen-binding portions, i.e., “antigen binding sites,” (e.g., fragments, subsequences, complementarity determining regions (CDRs)) that retain capacity to bind antigen, including (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR). Single chain antibodies are also included by reference in the term “antibody.”

Antibodies used in the immunoassays described herein preferably specifically bind to a kidney injury marker of the present invention. The term “specifically binds” is not intended to indicate that an antibody binds exclusively to its intended target since, as noted above, an antibody binds to any polypeptide displaying the epitope(s) to which the antibody binds. Rather, an antibody “specifically binds” if its affinity for its intended target is about 5-fold greater when compared to its affinity for a non-target molecule which does not display the appropriate epitope(s). Preferably the affinity of the antibody will be at least about 5 fold, preferably 10 fold, more preferably 25-fold, even more preferably 50-fold, and most preferably 100-fold or more, greater for a target molecule than its affinity for a non-target molecule. In preferred embodiments, Preferred antibodies bind with affinities of at least about 10⁷ M⁻¹, and preferably between about 10⁸ M⁻¹ to about 10⁹ M⁻¹, about 10⁹ M⁻¹ to about 10¹⁰ M⁻¹, or about 10¹⁰ M⁻¹ m to about 10¹² M⁻¹.

Affinity is calculated as K_(d)=k_(off)/k_(on) (k_(off) is the dissociation rate constant, K_(on) is the association rate constant and K_(d) is the equilibrium constant). Affinity can be determined at equilibrium by measuring the fraction bound (r) of labeled ligand at various concentrations (c). The data are graphed using the Scatchard equation: r/c=K(n−r): where r=moles of bound ligand/mole of receptor at equilibrium; c=free ligand concentration at equilibrium; K=equilibrium association constant; and n=number of ligand binding sites per receptor molecule. By graphical analysis, r/c is plotted on the Y-axis versus r on the X-axis, thus producing a Scatchard plot. Antibody affinity measurement by Scatchard analysis is well known in the art. See, e.g., van Erp et al., J. Immunoassay 12: 425-43, 1991; Nelson and Griswold, Comput. Methods Programs Biomed. 27: 65-8, 1988.

The term “epitope” refers to an antigenic determinant capable of specific binding to an antibody. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. Conformational and nonconformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents.

Numerous publications discuss the use of phage display technology to produce and screen libraries of polypeptides for binding to a selected analyte. See, e.g, Cwirla et al., Proc. Natl. Acad. Sci. USA 87, 6378-82, 1990; Devlin et al., Science 249, 404-6, 1990, Scott and Smith, Science 249, 386-88, 1990; and Ladner et al., U.S. Pat. No. 5,571,698. A basic concept of phage display methods is the establishment of a physical association between DNA encoding a polypeptide to be screened and the polypeptide. This physical association is provided by the phage particle, which displays a polypeptide as part of a capsid enclosing the phage genome which encodes the polypeptide. The establishment of a physical association between polypeptides and their genetic material allows simultaneous mass screening of very large numbers of phage bearing different polypeptides. Phage displaying a polypeptide with affinity to a target bind to the target and these phage are enriched by affinity screening to the target. The identity of polypeptides displayed from these phage can be determined from their respective genomes. Using these methods a polypeptide identified as having a binding affinity for a desired target can then be synthesized in bulk by conventional means. See, e.g., U.S. Pat. No. 6,057,098, which is hereby incorporated in its entirety, including all tables, FIGURES, and claims.

The antibodies that are generated by these methods may then be selected by first screening for affinity and specificity with the purified polypeptide of interest and, if required, comparing the results to the affinity and specificity of the antibodies with polypeptides that are desired to be excluded from binding. The screening procedure can involve immobilization of the purified polypeptides in separate wells of microtiter plates. The solution containing a potential antibody or groups of antibodies is then placed into the respective microtiter wells and incubated for about 30 min to 2 h. The microtiter wells are then washed and a labeled secondary antibody (for example, an anti-mouse antibody conjugated to alkaline phosphatase if the raised antibodies are mouse antibodies) is added to the wells and incubated for about 30 min and then washed. Substrate is added to the wells and a color reaction will appear where antibody to the immobilized polypeptide(s) are present.

The antibodies so identified may then be further analyzed for affinity and specificity in the assay design selected. In the development of immunoassays for a target protein, the purified target protein acts as a standard with which to judge the sensitivity and specificity of the immunoassay using the antibodies that have been selected. Because the binding affinity of various antibodies may differ; certain antibody pairs (e.g., in sandwich assays) may interfere with one another sterically, etc., assay performance of an antibody may be a more important measure than absolute affinity and specificity of an antibody.

Assay Correlations

The term “correlating” as used herein in reference to the use of biomarkers refers to comparing the presence or amount of the biomarker(s) in a patient to its presence or amount in persons known to suffer from, or known to be at risk of, a given condition; or in persons known to be free of a given condition. Often, this takes the form of comparing an assay result in the form of a biomarker concentration to a predetermined threshold selected to be indicative of the occurrence or nonoccurrence of a disease or the likelihood of some future outcome.

Selecting a diagnostic threshold involves, among other things, consideration of the probability of disease, distribution of true and false diagnoses at different test thresholds, and estimates of the consequences of treatment (or a failure to treat) based on the diagnosis. For example, when considering administering a specific therapy which is highly efficacious and has a low level of risk, few tests are needed because clinicians can accept substantial diagnostic uncertainty. On the other hand, in situations where treatment options are less effective and more risky, clinicians often need a higher degree of diagnostic certainty. Thus, cost/benefit analysis is involved in selecting a diagnostic threshold.

Suitable thresholds may be determined in a variety of ways. For example, one recommended diagnostic threshold for the diagnosis of acute myocardial infarction using cardiac troponin is the 97.5^(th) percentile of the concentration seen in a normal population. Another method may be to look at serial samples from the same patient, where a prior “baseline” result is used to monitor for temporal changes in a biomarker level.

Population studies may also be used to select a decision threshold. Receiver Operating Characteristic (“ROC”) arose from the field of signal detection theory developed during World War II for the analysis of radar images, and ROC analysis is often used to select a threshold able to best distinguish a “diseased” subpopulation from a “nondiseased” subpopulation. A false positive in this case occurs when the person tests positive, but actually does not have the disease. A false negative, on the other hand, occurs when the person tests negative, suggesting they are healthy, when they actually do have the disease. To draw a ROC curve, the true positive rate (TPR) and false positive rate (FPR) are determined as the decision threshold is varied continuously. Since TPR is equivalent with sensitivity and FPR is equal to 1—specificity, the ROC graph is sometimes called the sensitivity vs (1—specificity) plot. A perfect test will have an area under the ROC curve of 1.0; a random test will have an area of 0.5. A threshold is selected to provide an acceptable level of specificity and sensitivity.

In this context, “diseased” is meant to refer to a population having one characteristic (the presence of a disease or condition or the occurrence of some outcome) and “nondiseased” is meant to refer to a population lacking the characteristic. While a single decision threshold is the simplest application of such a method, multiple decision thresholds may be used. For example, below a first threshold, the absence of disease may be assigned with relatively high confidence, and above a second threshold the presence of disease may also be assigned with relatively high confidence. Between the two thresholds may be considered indeterminate. This is meant to be exemplary in nature only.

In addition to threshold comparisons, other methods for correlating assay results to a patient classification (occurrence or nonoccurrence of disease, likelihood of an outcome, etc.) include decision trees, rule sets, Bayesian methods, and neural network methods. These methods can produce probability values representing the degree to which a subject belongs to one classification out of a plurality of classifications.

Measures of test accuracy may be obtained as described in Fischer et al., Intensive Care Med. 29: 1043-51, 2003, and used to determine the effectiveness of a given biomarker. These measures include sensitivity and specificity, predictive values, likelihood ratios, diagnostic odds ratios, and ROC curve areas. The area under the curve (“AUC”) of a ROC plot is equal to the probability that a classifier will rank a randomly chosen positive instance higher than a randomly chosen negative one. The area under the ROC curve may be thought of as equivalent to the Mann-Whitney U test, which tests for the median difference between scores obtained in the two groups considered if the groups are of continuous data, or to the Wilcoxon test of ranks.

As discussed above, suitable tests may exhibit one or more of the following results on these various measures: a specificity of greater than 0.5, preferably at least 0.6, more preferably at least 0.7, still more preferably at least 0.8, even more preferably at least 0.9 and most preferably at least 0.95, with a corresponding sensitivity greater than 0.2, preferably greater than 0.3, more preferably greater than 0.4, still more preferably at least 0.5, even more preferably 0.6, yet more preferably greater than 0.7, still more preferably greater than 0.8, more preferably greater than 0.9, and most preferably greater than 0.95; a sensitivity of greater than 0.5, preferably at least 0.6, more preferably at least 0.7, still more preferably at least 0.8, even more preferably at least 0.9 and most preferably at least 0.95, with a corresponding specificity greater than 0.2, preferably greater than 0.3, more preferably greater than 0.4, still more preferably at least 0.5, even more preferably 0.6, yet more preferably greater than 0.7, still more preferably greater than 0.8, more preferably greater than 0.9, and most preferably greater than 0.95; at least 75% sensitivity, combined with at least 75% specificity; a ROC curve area of greater than 0.5, preferably at least 0.6, more preferably 0.7, still more preferably at least 0.8, even more preferably at least 0.9, and most preferably at least 0.95; an odds ratio different from 1, preferably at least about 2 or more or about 0.5 or less, more preferably at least about 3 or more or about 0.33 or less, still more preferably at least about 4 or more or about 0.25 or less, even more preferably at least about 5 or more or about 0.2 or less, and most preferably at least about 10 or more or about 0.1 or less; a positive likelihood ratio (calculated as sensitivity/(1-specificity)) of greater than 1, at least 2, more preferably at least 3, still more preferably at least 5, and most preferably at least 10; and or a negative likelihood ratio (calculated as (1-sensitivity)/specificity) of less than 1, less than or equal to 0.5, more preferably less than or equal to 0.3, and most preferably less than or equal to 0.1

Additional clinical indicia may be combined with the kidney injury marker assay result(s) of the present invention. These include other biomarkers related to renal status. Examples include the following, which recite the common biomarker name, followed by the Swiss-Prot entry number for that biomarker or its parent: Actin (P68133); Adenosine deaminase binding protein (DPP4, P27487); Alpha-1-acid glycoprotein 1 (P02763); Alpha-1-microglobulin (P02760); Albumin (P02768); Angiotensinogenase (Renin, P00797); Annexin A2 (P07355); Beta-glucuronidase (P08236); B-2-microglobulin (P61769); Beta-galactosidase (P16278); BMP-7 (P18075); Brain natriuretic peptide (proBNP, BNP-32, NTproBNP; P16860); Calcium-binding protein Beta (S100-beta, P04271); Carbonic anhydrase 9 (Q16790); Casein Kinase 2 (P68400); Clusterin (P10909); Complement C3 (P01024); Cysteine-rich protein (CYR61, 000622); Cytochrome C (P99999); Epidermal growth factor (EGF, P01133); Endothelin-1 (P05305); Exosomal Fetuin-A (P02765); Fatty acid-binding protein, heart (FABP3, P05413); Fatty acid-binding protein, liver (P07148); Ferritin (light chain, P02792; heavy chain P02794); Fructose-1,6-bisphosphatase (P09467); GRO-alpha (CXCL1, (P09341); Growth Hormone (P01241); Hepatocyte growth factor (P14210); Insulin-like growth factor I (P05019); Immunoglobulin G; Immunoglobulin Light Chains (Kappa and Lambda); Interferon gamma (P01579); Lysozyme (P61626); Interleukin-1 alpha (P01583); Interleukin-2 (P60568); Interleukin-4 (P05112); Interleukin-9 (P15248); Interleukin-12p40 (P29460); Interleukin-13 (P35225); Interleukin-16 (Q14005); L1 cell adhesion molecule (P32004); Lactate dehydrogenase (P00338); Leucine Aminopeptidase (P28838); Meprin A-alpha subunit (Q16819); Meprin A-beta subunit (Q16820); Midkine (P21741); MIP2-alpha (CXCL2, P19875); MMP-2 (P08253); MMP-9 (P14780); Netrin-1 (O95631); Neutral endopeptidase (P08473); Osteopontin (P10451); Renal papillary antigen 1 (RPA1); Renal papillary antigen 2 (RPA2); Retinol binding protein (P09455); Ribonuclease; S100 calcium-binding protein A6 (P06703); Serum Amyloid P Component (P02743); Sodium/Hydrogen exchanger isoform (NHE3, P48764); Spermidine/spermine N1-acetyltransferase (P21673); TGF-Beta1 (P01137); Transferrin (P02787); Trefoil factor 3 (TFF3, Q07654); Toll-Like protein 4 (O00206); Total protein; Tubulointerstitial nephritis antigen (Q9UJW2); Uromodulin (Tamm-Horsfall protein, P07911).

For purposes of risk stratification, Adiponectin (Q15848); Alkaline phosphatase (P05186); Aminopeptidase N (P15144); CalbindinD28k (P05937); Cystatin C (P01034); 8 subunit of F1FO ATPase (P03928); Gamma-glutamyltransferase (P19440); GSTa (alpha-glutathione-S-transferase, P08263); GSTpi (Glutathione-S-transferase P; GST class-pi; P09211); IGFBP-1 (P08833); IGFBP-2 (P18065); IGFBP-6 (P24592); Integral membrane protein 1 (Itm1, P46977); Interleukin-6 (P05231); Interleukin-8 (P10145); Interleukin-18 (Q14116); IP-10 (10 kDa interferon-gamma-induced protein, P02778); IRPR (IFRD1, O00458); Isovaleryl-CoA dehydrogenase (IVD, P26440); I-TAC/CXCL11 (O14625); Keratin 19 (P08727); Kim-1 (Hepatitis A virus cellular receptor 1, Q96D42); L-arginine:glycine amidinotransferase (P50440); Leptin (P41159); Lipocalin2 (NGAL, P80188); MCP-1 (P13500); MIG (Gamma-interferon-induced monokine Q07325); MIP-la (P10147); MIP-3a (P78556); MIP-1 beta (P13236); MIP-1d (Q16663); NAG (N-acetyl-beta-D-glucosaminidase, P54802); Organic ion transporter (OCT2, O15244); Osteoprotegerin (O00300); P8 protein (O60356); Plasminogen activator inhibitor 1 (PAI-1, P05121); ProANP (1-98) (P01160); Protein phosphatase 1-beta (PPI-beta, P62140); Rab GDI-beta (P50395); Renal kallikrein (P06870); RT1.B-1 (alpha) chain of the integral membrane protein (Q5Y7A8); soluble tumor necrosis factor receptor superfamily member 1A (sTNFR-I, P19438); soluble tumor necrosis factor receptor superfamily member 1B (sTNFR-II, P20333); Tissue inhibitor of metalloproteinases 3 (TIMP-3, P35625); uPAR (Q03405) may be combined with the kidney injury marker assay result(s) of the present invention.

Other clinical indicia which may be combined with the kidney injury marker assay result(s) of the present invention includes demographic information (e.g., weight, sex, age, race), medical history (e.g., family history, type of surgery, pre-existing disease such as aneurism, congestive heart failure, preeclampsia, eclampsia, diabetes mellitus, hypertension, coronary artery disease, proteinuria, renal insufficiency, or sepsis, type of toxin exposure such as NSAIDs, cyclosporines, tacrolimus, aminoglycosides, foscarnet, ethylene glycol, hemoglobin, myoglobin, ifosfamide, heavy metals, methotrexate, radiopaque contrast agents, or streptozotocin), clinical variables (e.g., blood pressure, temperature, respiration rate), risk scores (APACHE score, PREDICT score, TIMI Risk Score for UA/NSTEMI, Framingham Risk Score), a urine total protein measurement, a glomerular filtration rate, an estimated glomerular filtration rate, a urine production rate, a serum or plasma creatinine concentration, a renal papillary antigen 1 (RPA1) measurement; a renal papillary antigen 2 (RPA2) measurement; a urine creatinine concentration, a fractional excretion of sodium, a urine sodium concentration, a urine creatinine to serum or plasma creatinine ratio, a urine specific gravity, a urine osmolality, a urine urea nitrogen to plasma urea nitrogen ratio, a plasma BUN to creatinine ratio, and/or a renal failure index calculated as urine sodium/(urine creatinine/plasma creatinine). Other measures of renal function which may be combined with the kidney injury marker assay result(s) are described hereinafter and in Harrison's Principles of Internal Medicine, 17^(th) Ed., McGraw Hill, New York, pages 1741-1830, and Current Medical Diagnosis & Treatment 2008, 47^(th) Ed, McGraw Hill, New York, pages 785-815, each of which are hereby incorporated by reference in their entirety.

Combining assay results/clinical indicia in this manner can comprise the use of multivariate logistical regression, loglinear modeling, neural network analysis, n-of-m analysis, decision tree analysis, etc. This list is not meant to be limiting.

Diagnosis of Acute Renal Failure

As noted above, the terms “acute renal (or kidney) injury” and “acute renal (or kidney) failure” as used herein are defined in part in terms of changes in serum creatinine from a baseline value. Most definitions of ARF have common elements, including the use of serum creatinine and, often, urine output. Patients may present with renal dysfunction without an available baseline measure of renal function for use in this comparison. In such an event, one may estimate a baseline serum creatinine value by assuming the patient initially had a normal GFR. Glomerular filtration rate (GFR) is the volume of fluid filtered from the renal (kidney) glomerular capillaries into the Bowman's capsule per unit time. Glomerular filtration rate (GFR) can be calculated by measuring any chemical that has a steady level in the blood, and is freely filtered but neither reabsorbed nor secreted by the kidneys. GFR is typically expressed in units of ml/min:

${GFR} = \frac{{Urine}{Concentration} \times {Urine}{Flow}}{{Plasma}{Concentration}}$

By normalizing the GFR to the body surface area, a GFR of approximately 75-100 ml/min per 1.73 m² can be assumed. The rate therefore measured is the quantity of the substance in the urine that originated from a calculable volume of blood.

There are several different techniques used to calculate or estimate the glomerular filtration rate (GFR or eGFR). In clinical practice, however, creatinine clearance is used to measure GFR. Creatinine is produced naturally by the body (creatinine is a metabolite of creatine, which is found in muscle). It is freely filtered by the glomerulus, but also actively secreted by the renal tubules in very small amounts such that creatinine clearance overestimates actual GFR by 10-20%. This margin of error is acceptable considering the ease with which creatinine clearance is measured.

Creatinine clearance (CCr) can be calculated if values for creatinine's urine concentration (U_(Cr)), urine flow rate (V), and creatinine's plasma concentration (P_(Cr)) are known. Since the product of urine concentration and urine flow rate yields creatinine's excretion rate, creatinine clearance is also said to be its excretion rate (U_(Cr)×V) divided by its plasma concentration. This is commonly represented mathematically as:

$C_{Cr} = \frac{U_{Cr} \times V}{P_{Cr}}$

Commonly a 24 hour urine collection is undertaken, from empty-bladder one morning to the contents of the bladder the following morning, with a comparative blood test then taken:

$C_{Cr} = \frac{U_{Cr} \times 24 - {hour}{volume}}{P_{Cr} \times 24 \times 60{mins}}$

To allow comparison of results between people of different sizes, the CCr is often corrected for the body surface area (BSA) and expressed compared to the average sized man as ml/min/1.73 m2. While most adults have a BSA that approaches 1.7 (1.6-1.9), extremely obese or slim patients should have their CCr corrected for their actual BSA:

$C_{{Cr} - {corrected}} = \frac{C_{Cr} \times 1.73}{BSA}$

The accuracy of a creatinine clearance measurement (even when collection is complete) is limited because as glomerular filtration rate (GFR) falls creatinine secretion is increased, and thus the rise in serum creatinine is less. Thus, creatinine excretion is much greater than the filtered load, resulting in a potentially large overestimation of the GFR (as much as a twofold difference). However, for clinical purposes it is important to determine whether renal function is stable or getting worse or better. This is often determined by monitoring serum creatinine alone. Like creatinine clearance, the serum creatinine will not be an accurate reflection of GFR in the non-steady-state condition of ARF. Nonetheless, the degree to which serum creatinine changes from baseline will reflect the change in GFR. Serum creatinine is readily and easily measured and it is specific for renal function.

For purposes of determining urine output on a Urine output on a mL/kg/hr basis, hourly urine collection and measurement is adequate. In the case where, for example, only a cumulative 24-h output was available and no patient weights are provided, minor modifications of the RIFLE urine output criteria have been described. For example, Bagshaw et al., Nephrol. Dial. Transplant. 23: 1203-1210, 2008, assumes an average patient weight of 70 kg, and patients are assigned a RIFLE classification based on the following: <35 mL/h (Risk), <21 mL/h (Injury) or <4 mL/h (Failure).

Selecting a Treatment Regimen

Once a diagnosis is obtained, the clinician can readily select a treatment regimen that is compatible with the diagnosis, such as initiating renal replacement therapy, withdrawing delivery of compounds that are known to be damaging to the kidney, kidney transplantation, delaying or avoiding procedures that are known to be damaging to the kidney, modifying diuretic administration, initiating goal directed therapy, etc. The skilled artisan is aware of appropriate treatments for numerous diseases discussed in relation to the methods of diagnosis described herein. See, e.g., Merck Manual of Diagnosis and Therapy, 17th Ed. Merck Research Laboratories, Whitehouse Station, N J, 1999. In addition, since the methods and compositions described herein provide prognostic information, the markers of the present invention may be used to monitor a course of treatment. For example, improved or worsened prognostic state may indicate that a particular treatment is or is not efficacious.

One skilled in the art readily appreciates that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The examples provided herein are representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention.

Example 1: Contrast-Induced Nephropathy Sample Collection

The objective of this sample collection study is to collect samples of plasma and urine and clinical data from patients before and after receiving intravascular contrast media. Approximately 250 adults undergoing radiographic/angiographic procedures involving intravascular administration of iodinated contrast media are enrolled. To be enrolled in the study, each patient must meet all of the following inclusion criteria and none of the following exclusion criteria:

Inclusion Criteria

males and females 18 years of age or older; undergoing a radiographic/angiographic procedure (such as a CT scan or coronary intervention) involving the intravascular administration of contrast media; expected to be hospitalized for at least 48 hours after contrast administration. able and willing to provide written informed consent for study participation and to comply with all study procedures.

Exclusion Criteria

renal transplant recipients; acutely worsening renal function prior to the contrast procedure; already receiving dialysis (either acute or chronic) or in imminent need of dialysis at enrollment; expected to undergo a major surgical procedure (such as involving cardiopulmonary bypass) or an additional imaging procedure with contrast media with significant risk for further renal insult within the 48 hrs following contrast administration; participation in an interventional clinical study with an experimental therapy within the previous 30 days; known infection with human immunodeficiency virus (HIV) or a hepatitis virus.

Immediately prior to the first contrast administration (and after any pre-procedure hydration), an EDTA anti-coagulated blood sample (10 mL) and a urine sample (10 mL) are collected from each patient. Blood and urine samples are then collected at 4 (±0.5), 8 (±1), 24 (±2) 48 (±2), and 72 (±2) hrs following the last administration of contrast media during the index contrast procedure. Blood is collected via direct venipuncture or via other available venous access, such as an existing femoral sheath, central venous line, peripheral intravenous line or hep-lock. These study blood samples are processed to plasma at the clinical site, frozen and shipped to Astute Medical, Inc., San Diego, Calif. The study urine samples are frozen and shipped to Astute Medical, Inc.

Serum creatinine is assessed at the site immediately prior to the first contrast administration (after any pre-procedure hydration) and at 4 (±0.5), 8 (±1), 24 (±2) and 48 (±2)), and 72 (±2) hours following the last administration of contrast (ideally at the same time as the study samples are obtained). In addition, each patient's status is evaluated through day 30 with regard to additional serum and urine creatinine measurements, a need for dialysis, hospitalization status, and adverse clinical outcomes (including mortality).

Prior to contrast administration, each patient is assigned a risk based on the following assessment: systolic blood pressure <80 mm Hg=5 points; intra-arterial balloon pump=5 points; congestive heart failure (Class III-IV or history of pulmonary edema)=5 points; age >75 yrs=4 points; hematocrit level <39% for men, <35% for women=3 points; diabetes=3 points; contrast media volume=1 point for each 100 mL; serum creatinine level >1.5 g/dL=4 points OR estimated GFR 40-60 mL/min/1.73 m²=2 points, 20-40 mL/min/1.73 m²=4 points, <20 mL/min/1.73 m²=6 points. The risks assigned are as follows: risk for CIN and dialysis: 5 or less total points=risk of CIN—7.5%, risk of dialysis—0.04%; 6-10 total points=risk of CIN—14%, risk of dialysis—0.12%; 11-16 total points=risk of CIN—26.1%, risk of dialysis—1.09%; >16 total points=risk of CIN—57.3%, risk of dialysis—12.8%.

Example 2: Cardiac Surgery Sample Collection

The objective of this sample collection study is to collect samples of plasma and urine and clinical data from patients before and after undergoing cardiovascular surgery, a procedure known to be potentially damaging to kidney function. Approximately 900 adults undergoing such surgery are enrolled. To be enrolled in the study, each patient must meet all of the following inclusion criteria and none of the following exclusion criteria:

Inclusion Criteria

males and females 18 years of age or older; undergoing cardiovascular surgery; Toronto/Ottawa Predictive Risk Index for Renal Replacement risk score of at least 2 (Wijeysundera et al., JAMA 297: 1801-9, 2007); and able and willing to provide written informed consent for study participation and to comply with all study procedures.

Exclusion Criteria

known pregnancy; previous renal transplantation; acutely worsening renal function prior to enrollment (e.g., any category of RIFLE criteria); already receiving dialysis (either acute or chronic) or in imminent need of dialysis at enrollment; currently enrolled in another clinical study or expected to be enrolled in another clinical study within 7 days of cardiac surgery that involves drug infusion or a therapeutic intervention for AKI; known infection with human immunodeficiency virus (HIV) or a hepatitis virus.

Within 3 hours prior to the first incision (and after any pre-procedure hydration), an EDTA anti-coagulated blood sample (10 mL), whole blood (3 mL), and a urine sample (35 mL) are collected from each patient. Blood and urine samples are then collected at 3 (±0.5), 6 (±0.5), 12 (±1), 24 (±2) and 48 (±2) hrs following the procedure and then daily on days 3 through 7 if the subject remains in the hospital. Blood is collected via direct venipuncture or via other available venous access, such as an existing femoral sheath, central venous line, peripheral intravenous line or hep-lock. These study blood samples are frozen and shipped to Astute Medical, Inc., San Diego, Calif. The study urine samples are frozen and shipped to Astute Medical, Inc.

Example 3: Acutely Ill Subject Sample Collection

The objective of this study is to collect samples from acutely ill patients. Approximately 2200 adults expected to be in the ICU for at least 48 hours will be enrolled. To be enrolled in the study, each patient must meet all of the following inclusion criteria and none of the following exclusion criteria:

Inclusion Criteria

males and females 18 years of age or older; Study population 1: approximately 300 patients that have at least one of: shock (SBP<90 mmHg and/or need for vasopressor support to maintain MAP>60 mmHg and/or documented drop in SBP of at least 40 mmHg); and sepsis; Study population 2: approximately 300 patients that have at least one of: IV antibiotics ordered in computerized physician order entry (CPOE) within 24 hours of enrollment; contrast media exposure within 24 hours of enrollment; increased Intra-Abdominal Pressure with acute decompensated heart failure; and severe trauma as the primary reason for ICU admission and likely to be hospitalized in the ICU for 48 hours after enrollment; Study population 3: approximately 300 patients expected to be hospitalized through acute care setting (ICU or ED) with a known risk factor for acute renal injury (e.g. sepsis, hypotension/shock (Shock=systolic BP<90 mmHg and/or the need for vasopressor support to maintain a MAP>60 mmHg and/or a documented drop in SBP>40 mmHg), major trauma, hemorrhage, or major surgery); and/or expected to be hospitalized to the ICU for at least 24 hours after enrollment;

Study population 4: approximately 1000 patients that are 21 years of age or older, within 24 hours of being admitted into the ICU, expected to have an indwelling urinary catheter for at least 48 hours after enrollment, and have at least one of the following acute conditions within 24 hours prior to enrollment:

(i) respiratory SOFA score of ≥2 (PaO2/FiO2<300), (ii) cardiovascular SOFA score of ≥1 (MAP<70 mm Hg and/or any vasopressor required). Study population 5: approximately 300 patients that are 21 years of age or older, receiving care in the ICU, have an indwelling urinary catheter as standard care at the time of enrollment, have acute kidney injury (KDIGO stage 2 or stage 3) at the time of the first sample collection, and have their first sample collected within 36 hours of meeting KDIGO stage 2 criteria

Exclusion Criteria

known pregnancy; prisoners or institutionalized individuals; previous renal transplantation; comfort-measures-only status for study population 5; known acutely worsening renal function prior to enrollment (e.g., any category of RIFLE criteria) for study populations 1, 2, 3, and 4; received dialysis (either acute or chronic) within 5 days prior to enrollment or in imminent need of dialysis at the time of enrollment; known infection with human immunodeficiency virus (HIV) or a hepatitis virus; meets any of the following in study populations 4 and 5:

-   -   (i) active bleeding with an anticipated need for >4 units PRBC         in a day;     -   (ii) hemoglobin <7 g/dL;     -   (iii) any other condition that in the physician's opinion would         contraindicate drawing serial blood samples for clinical study         purposes;         meets only the SBP<90 mmHg inclusion criterion set forth above,         and does not have shock in the attending physician's or         principal investigator's opinion.

After obtaining informed consent, an EDTA anti-coagulated blood sample (10 mL) and a urine sample (25-50 mL) are collected from each patient. Blood and urine samples are then collected at 4 (±0.5) and 8 (±1) hours after contrast administration (if applicable); at 12 (±1), 24 (±2), 36 (±2), 48 (±2), 60 (±2), 72 (±2), and 84 (±2) hours after enrollment, and thereafter daily up to day 7 to day 14 while the subject is hospitalized. Blood is collected via direct venipuncture or via other available venous access, such as an existing femoral sheath, central venous line, peripheral intravenous line or hep-lock. These study blood samples are processed to plasma at the clinical site, frozen and shipped to Astute Medical, Inc., San Diego, Calif. The study urine samples are frozen and shipped to Astute Medical, Inc.

Example 4. Immunoassay Format

Analytes are measured using standard sandwich enzyme immunoassay techniques. A first antibody which binds the analyte is immobilized in wells of a 96 well polystyrene microplate. Analyte standards and test samples are pipetted into the appropriate wells and any analyte present is bound by the immobilized antibody. After washing away any unbound substances, a horseradish peroxidase-conjugated second antibody which binds the analyte is added to the wells, thereby forming sandwich complexes with the analyte (if present) and the first antibody. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution comprising tetramethylbenzidine and hydrogen peroxide is added to the wells. Color develops in proportion to the amount of analyte present in the sample. The color development is stopped and the intensity of the color is measured at 540 nm or 570 nm. An analyte concentration is assigned to the test sample by comparison to a standard curve determined from the analyte standards.

Table of Swiss-Prot numbers, preferred names and units for the analytes in the examples below.

Swiss-Prot Number Preferred Name Units P01033 Metalloproteinase inhibitor 1 pg/ml P16035 Metalloproteinase inhibitor 2 pg/ml Q99727 Metalloproteinase inhibitor 4 pg/ml Q16663 C—C motif chemokine 15 pg/ml P55774 C—C motif chemokine 18 ng/ml P55773 C—C motif chemokine 23 ng/ml O00175 C—C motif chemokine 24 pg/ml

Example 5. Apparently Healthy Donor and Chronic Disease Patient Samples

Human urine samples from donors with no known chronic or acute disease (“Apparently Healthy Donors”) were purchased from two vendors (Golden West Biologicals, Inc., 27625 Commerce Center Dr., Temecula, Calif. 92590 and Virginia Medical Research, Inc., 915 First Colonial Rd., Virginia Beach, Va. 23454). The urine samples were shipped and stored frozen at less than −20° C. The vendors supplied demographic information for the individual donors including gender, race (Black/White), smoking status and age.

Human urine samples from donors with various chronic diseases (“Chronic Disease Patients”) including congestive heart failure, coronary artery disease, chronic kidney disease, chronic obstructive pulmonary disease, diabetes mellitus and hypertension were purchased from Virginia Medical Research, Inc., 915 First Colonial Rd., Virginia Beach, Va. 23454. The urine samples were shipped and stored frozen at less than −20 degrees centigrade. The vendor provided a case report form for each individual donor with age, gender, race (Black/White), smoking status and alcohol use, height, weight, chronic disease(s) diagnosis, current medications and previous surgeries.

Example 6. Use of Metalloproteinase Inhibitor 1 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 1 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 6.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 5950 17800 5970 17700 5950 17300 Average 9410 33100 8890 32200 8800 31800 Stdev 9010 35200 7930 34800 8170 34600 p (t-test) 8.0E−7 8.9E−6 3.1E−5 Min 634 297 634 297 634 297 Max 39400 146000 32600 146000 32600 146000 n (Patient) 57 260 46 271 41 276 sCr only Median 7940 18500 7940 18100 7940 18100 Average 12800 33700 13000 33500 11700 33500 Stdev 16300 35700 16400 35600 11300 35800 p (t-test) 1.3E−6 2.7E−6 8.7E−7 Min 634 297 634 297 634 297 Max 116000 146000 116000 146000 65500 146000 n (Patient) 76 240 74 242 70 246 UO only Median 10800 21300 9980 20500 9520 20800 Average 23500 37700 23000 36200 22100 35700 Stdev 30800 35700 30600 35400 29800 35400 p (t-test) 2.2E−4 4.4E−4 2.7E−4 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 127000 146000 n (Patient) 194 121 173 142 156 159 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.75 0.70 0.66 0.75 0.69 0.65 0.75 0.70 0.66 SE 0.031 0.032 0.032 0.033 0.032 0.031 0.035 0.032 0.030 p Value 2.2E−16 2.7E−10 7.5E−7 6.6E−14 2.2E−9 8.9E−7 2.7E−13 8.0E−10 1.2E−7 nCohort Recovered 57 76 194 46 74 173 41 70 156 nCohort Non- 260 240 121 271 242 142 276 246 159 recovered Cutoff Quartile 2 6030 6020 6080 6030 6020 6080 6030 6020 6080 Sensitivity 82% 81% 88% 80% 81% 86% 79% 80% 85% Specificity 54% 43% 34% 52% 43% 34% 54% 43% 35% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity 58% 58% 66% 56% 58% 63% 56% 58% 63% Specificity 84% 76% 60% 87% 76% 61% 88% 77% 63% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity 30% 31% 35% 30% 31% 33% 29% 31% 33% Specificity 98% 95% 81% 100% 95% 82% 100% 96% 83% OR Quartile 2 5.27 3.24 3.85 4.28 3.16 3.16 4.45 3.02 3.06 p Value 8.6E−8 3.5E−5 2.9E−5 LIE−5 5.5E−5 7.2E−5 1.7E−5 1.4E−4 5.6E−5 Lower limit of 95% 2.87 1.86 2.05 2.24 1.81 1.79 2.25 1.71 1.78 CI Upper limit of 95% 9.67 5.64 7.24 8.21 5.53 5.57 8.78 5.32 5.28 CI OR Quartile 3 7.27 4.51 2.90 8.64 4.27 2.67 9.09 4.61 2.86 p Value 2.4E−7 5.1E−7 1.0E−5 2.1E−6 1.4E−6 2.6E−5 7.4E−6 1.0E−6 6.4E−6 Lower limit of 95% 3.42 2.51 1.81 3.55 2.37 1.69 3.46 2.50 1.81 CI Upper limit of 95% 15.4 8.12 4.66 21.1 7.70 4.22 23.9 8.50 4.52 CI OR Quartile 4 24.4 8.18 2.26 39.1 7.86 2.18 34.0 9.98 2.50 p Value 0.0017 7.8E−5 0.0021 0.010 1.1E−4 0.0033 0.014 1.5E−4 7.9E−4 Lower limit of 95% 3.32 2.88 1.34 2.38 2.77 1.30 2.07 3.04 1.46 CI Upper limit of 95% 180 23.2 3.79 642 22.3 3.66 559 32.7 4.27 CI

TABLE 6.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or scrum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 6460 20500 6980 19000 6510 18500 Average 13400 34900 14200 33700 14400 33000 Stdev 18900 35800 19900 35500 20800 35100 p (t-test) 1.2E−7 5.1E−6 2.7E−5 Min 634 297 634 297 634 297 Max 110000 146000 110000 146000 110000 146000 n (Patient) 90 227 79 238 71 246 sCr only Median 8310 20500 8310 20000 8210 19800 Average 15500 35500 15700 35200 14700 35200 Stdev 21000 36300 21100 36300 19000 36400 p (t-test) 2.9E−7 6.5E−7 2.4E−7 Min 634 297 634 297 634 297 Max 116000 146000 116000 146000 110000 146000 n (Patient) 106 211 104 213 99 218 UO only Median 11200 20500 11000 19200 11000 19300 Average 25400 36200 25900 33900 25000 34000 Stdev 31900 35500 32300 34700 31500 35300 p (t-test) 0.0071 0.039 0.017 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 127000 146000 n (Patient) 212 103 194 121 178 137 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.74 0.70 0.64 0.71 0.69 0.61 0.71 0.70 0.62 SE 0.029 0.030 0.034 0.031 0.030 0.033 0.032 0.030 0.032 p Value 2.2E−16 1.5E−11 5.3E−5 4.3E−12 1.2E−10 5.9E−4 1.8E−11 2.2E−11 2.6E−4 nCohort Recovered 90 106 212 79 104 194 71 99 178 nCohort Non- 227 211 103 238 213 121 246 218 137 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 85% 84% 90% 82% 84% 87% 82% 83% 85% Specificity 49% 42% 33% 47% 42% 32% 48% 42% 33% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 61% 62% 67% 59% 61% 61% 58% 61% 61% Specificity 78% 73% 58% 76% 72% 57% 77% 74% 58% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 32% 33% 31% 31% 33% 29% 30% 33% 31% Specificity 92% 91% 78% 91% 90% 77% 90% 91% 79% OR Quartile 2 5.25 3.84 4.49 4.11 3.73 3.16 4.10 3.60 2.67 p Value 3.1E−9 7.1E−7 3.7E−5 5.6E−7 1.2E−6 2.0E−4 1.0E−6 2.4E−6 6.0E−4 Lower limit of 95% 3.03 2.26 2.20 2.36 2.19 1.72 2.33 2.12 1.52 CI Upper limit of 95% 9.08 6.54 9.15 7.15 6.35 5.78 7.23 6.14 4.68 CI OR Quartile 3 5.53 4.26 2.80 4.51 4.05 2.06 4.77 4.39 2.11 p Value 2.8E−9 2.4E−8 4.1E−5 3.1E−7 7.4E−8 0.0022 5.5E−7 3.1E−8 0.0013 Lower limit of 95% 3.14 2.56 1.71 2.53 2.43 1.30 2.59 2.60 1.34 CI Upper limit of 95% 9.72 7.09 4.59 8.03 6.74 3.28 8.80 7.42 3.32 CI OR Quartile 4 5.62 4.77 1.58 4.55 4.60 1.39 3.86 4.83 1.68 p Value 3.7E−5 1.7E−5 0.089 3.1E−4 2.6E−5 0.21 0.0014 3.1E−5 0.046 Lower limit of 95% 2.48 2.34 0.933 2.00 2.26 0.827 1.69 2.30 1.01 CI Upper limit of 95% 12.8 9.71 2.68 10.4 9.38 2.33 8.82 10.1 2.81 CI

TABLE 6.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or scrum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 7170 20500 7180 20200 7150 19300 Average 15100 36500 15400 35700 15800 34600 Stdev 20200 36700 20700 36400 21500 36000 p (t-test) 1.9E−8 1.9E−7 2.6E−6 Min 475 297 475 297 475 297 Max 110000 146000 110000 146000 110000 146000 n (Patient) 114 203 107 210 97 220 sCr only Median 8610 20500 8810 20000 8610 19800 Average 16800 36900 17100 36400 16300 36300 Stdev 21800 37100 22000 37100 20400 37200 p (t-test) 8.2E−8 3.2E−7 1.3E−7 Min 475 297 475 297 475 297 Max 116000 146000 116000 146000 110000 146000 n (Patient) 127 190 124 193 119 198 UO only Median 11600 20500 11500 19400 11000 20500 Average 26000 35300 25900 34500 24700 35300 Stdev 32000 35500 32000 35400 31000 35900 p (t-test) 0.021 0.030 0.0057 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 127000 146000 n (Patient) 214 101 204 111 190 125 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.69 0.62 0.71 0.68 0.61 0.70 0.69 0.63 SE 0.028 0.029 0.035 0.029 0.030 0.034 0.030 0.030 0.033 p Value 1.6E−15 2.6E−11 6.7E−4 2.9E−13 9.0E−10 0.0013 6.9E−12 2.9E−10 1.2E−4 nCohort Recovered 114 127 214 107 124 204 97 119 190 nCohort Non- 203 190 101 210 193 111 220 198 125 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 87% 86% 87% 85% 85% 86% 84% 84% 85% Specificity 46% 41% 31% 45% 40% 31% 45% 40% 32% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 64% 63% 64% 62% 62% 62% 60% 62% 62% Specificity 74% 69% 57% 73% 69% 56% 73% 70% 58% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 34% 35% 30% 33% 34% 29% 31% 34% 32% Specificity 90% 89% 77% 90% 89% 77% 89% 89% 79% OR Quartile 2 5.47 4.19 3.02 4.70 3.82 2.65 4.39 3.64 2.57 p Value 1.2E−9 2.0E−7 8.7E−4 1.8E−8 8.4E−7 0.0016 7.5E−8 1.8E−6 0.0013 Lower limit of 95% 3.16 2.44 1.57 2.74 2.24 1.45 2.56 2.14 1.45 CI Upper limit of 95% 9.45 7.18 5.79 8.05 6.51 4.87 7.52 6.19 4.58 CI OR Quartile 3 4.88 3.87 2.35 4.37 3.58 2.12 4.17 3.78 2.28 p Value 7.9E−10 3.0E−8 6.1E−4 1.4E−8 1.7E−7 0.0018 9.2E−8 7.8E−8 4.7E−4 Lower limit of 95% 2.94 2.40 1.44 2.63 2.22 1.32 2.47 2.33 1.44 CI Upper limit of 95% 8.09 6.24 3.83 7.27 5.78 3.41 7.05 6.14 3.62 CI OR Quartile 4 4.82 4.30 1.42 4.27 4.08 1.35 3.57 4.17 1.82 p Value 7.0E−6 5.9E−6 0.19 3.5E−5 1.2E−5 0.26 2.9E−4 1.5E−5 0.022 Lower limit of 95% 2.43 2.29 0.835 2.15 2.17 0.801 1.79 2.18 1.09 CI Upper limit of 95% 9.58 8.07 2.42 8.49 7.67 2.29 7.12 7.96 3.05 CI

TABLE 6.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or scrum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 8180 22000 8150 21100 7940 20000 Average 15800 38300 16200 37000 16600 35700 Stdev 21300 37200 21900 36900 22700 36300 p (t-test) 1.1E−9 2.9E−8 6.2E−7 Min 419 297 419 297 419 297 Max 111000 146000 111000 146000 111000 146000 n (Patient) 134 183 125 192 114 203 sCr only Median 8710 21900 8740 21300 8730 21600 Average 17200 38000 17400 37700 16600 37900 Stdev 22900 37300 23000 37200 21600 37500 p (t-test) 1.6E−8 4.2E−8 9.3E−9 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 111000 146000 n (Patient) 140 177 138 179 135 182 UO only Median 11600 21300 11200 20300 11000 20500 Average 25600 36300 25600 35400 24600 35900 Stdev 31900 35600 31800 35600 30800 36300 p (t-test) 0.0076 0.013 0.0032 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 127000 146000 n (Patient) 216 99 207 108 194 121 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.71 0.63 0.72 0.70 0.62 0.71 0.71 0.63 SE 0.028 0.029 0.035 0.028 0.029 0.034 0.029 0.029 0.033 p Value 0 3.9E−13 1.9E−4 2.8E−14 4.1E−12 2.7E−4 6.0E−13 5.8E−13 6.7E−5 nCohort Recovered 134 140 216 125 138 207 114 135 194 nCohort Non- 183 177 99 192 179 108 203 182 121 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 88% 87% 88% 86% 87% 88% 85% 86% 86% Specificity 43% 40% 31% 42% 40% 32% 43% 40% 32% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 67% 66% 66% 64% 65% 63% 63% 65% 63% Specificity 72% 69% 57% 71% 69% 57% 72% 70% 58% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 37% 36% 33% 35% 36% 31% 33% 36% 33% Specificity 90% 89% 79% 90% 88% 78% 89% 89% 80% OR Quartile 2 5.42 4.46 3.26 4.70 4.28 3.42 4.35 4.19 2.87 p Value 3.8E−9 1.2E−7 5.3E−4 2.6E−8 2.1E−7 2.0E−4 7.9E−8 2.6E−7 5.1E−4 Lower limit of 95% 3.09 2.57 1.67 2.73 2.47 1.79 2.54 2.43 1.58 CI Upper limit of 95% 9.50 7.76 6.36 8.10 7.41 6.55 7.43 7.22 5.21 CI OR Quartile 3 5.24 4.29 2.53 4.41 4.07 2.21 4.28 4.23 2.31 p Value 2.8E−11 1.8E−9 2.4E−4 2.3E−9 6.2E−9 0.0011 1.0E−8 3.1E−9 4.4E−4 Lower limit of 95% 3.22 2.67 1.54 2.71 2.53 1.37 2.60 2.62 1.45 CI Upper limit of 95% 8.54 6.89 4.15 7.17 6.53 3.56 7.04 6.81 3.68 CI OR Quartile 4 5.38 4.39 1.85 4.62 4.24 1.65 3.83 4.44 1.96 p Value 3.4E−7 1.6E−6 0.023 3.5E−6 2.7E−6 0.060 4.8E−5 2.1E−6 0.011 Lower limit of 95% 2.82 2.40 1.09 2.42 2.32 0.980 2.00 2.40 1.17 CI Upper limit of 95% 10.3 8.03 3.14 8.81 7.76 2.79 7.31 8.23 3.29 CI

TABLE 6.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or scrum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 10600 20800 10000 20700 10000 20700 Average 22100 38700 22200 38200 22200 38200 Stdev 28300 37600 28500 37300 28500 37300 p (t-test) 9.9E−6 2.0E−5 2.0E−5 Min 297 399 297 399 297 399 Max 127000 146000 127000 146000 127000 146000 n (Patient) 188 129 185 132 185 132 sCr only Median 9400 22000 9350 22000 9330 22000 Average 18800 39100 18900 38600 18300 39000 Stdev 24400 37900 24600 37700 23400 38100 p (t-test) 3.0E−8 8.7E−8 1.5E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 111000 146000 n (Patient) 160 157 157 160 156 161 UO only Median 11300 21500 11600 20800 11300 21100 Average 24300 38300 24500 37500 24500 37300 Stdev 29900 38000 30100 37800 30200 37400 p (t-test) 4.4E−4 0.0010 0.0012 Min 297 399 297 399 297 399 Max 127000 146000 127000 146000 127000 146000 n (Patient) 211 104 208 107 205 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.70 0.64 0.66 0.69 0.63 0.66 0.70 0.63 SE 0.032 0.029 0.034 0.031 0.029 0.034 0.031 0.029 0.034 p Value 3.5E−7 1.9E−11 5.5E−5 3.6E−7 8.3E−11 1.1E−4 3.6E−7 1.5E−11 9.6E−5 nCohort Recovered 188 160 211 185 157 208 185 156 205 nCohort Non- 129 157 104 132 160 107 132 161 110 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 87% 88% 87% 87% 88% 87% 87% 88% 86% Specificity 33% 38% 31% 34% 38% 31% 34% 38% 31% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 66% 66% 65% 65% 66% 64% 65% 66% 64% Specificity 61% 66% 57% 61% 66% 57% 61% 66% 57% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 37% 37% 35% 36% 36% 34% 36% 37% 34% Specificity 83% 86% 80% 83% 86% 79% 83% 87% 80% OR Quartile 2 3.24 4.36 2.86 3.41 4.21 3.02 3.41 4.29 2.87 p Value 1.0E−4 5.6E−7 0.0012 5.1E−5 7.3E−7 6.3E−4 5.1E−5 5.4E−7 8.4E−4 Lower limit of 95% 1.79 2.45 1.52 1.88 2.39 1.60 1.88 2.43 1.55 CI Upper limit of 95% 5.87 7.76 5.40 6.17 7.44 5.69 6.17 7.58 5.34 CI OR Quartile 3 2.98 3.75 2.54 2.87 3.64 2.29 2.87 3.75 2.33 p Value 4.7E−6 2.5E−8 1.8E−4 8.5E−6 4.6E−8 7.3E−4 8.5E−6 2.5E−8 5.2E−4 Lower limit of 95% 1.87 2.35 1.56 1.80 2.29 1.41 1.80 2.35 1.44 CI Upper limit of 95% 4.75 5.96 4.13 4.56 5.79 3.69 4.56 5.96 3.75 CI OR Quartile 4 2.89 3.67 2.07 2.73 3.49 1.95 2.73 3.72 1.97 p Value 6.7E−5 4.2E−6 0.0066 1.5E−4 9.8E−6 0.013 1.5E−4 4.4E−6 0.011 Lower limit of 95% 1.71 2.11 1.22 1.62 2.01 1.15 1.62 2.12 1.17 CI Upper limit of 95% 4.87 6.40 3.50 4.60 6.07 3.28 4.60 6.51 3.31 CI

TABLE 6.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 253000 499000 245000 494000 245000 472000 Average 314000 530000 280000 527000 285000 523000 Stdev 213000 274000 171000 275000 178000 275000 p (t-test) 1.5E−7 3.4E−8 3.8E−7 Min 66700 13600 66700 13600 66700 13600 Max 975000 1120000 872000 1120000 872000 1120000 n (Patient) 52 275 42 285 38 289 sCr only Median 257000 519000 257000 519000 257000 511000 Average 341000 535000 333000 535000 341000 530000 Stdev 237000 270000 231000 270000 236000 271000 p (t-test) 1.1E−7 4.6E−8 5.7E−7 Min 66700 13600 66700 13600 66700 13600 Max 950000 1120000 950000 1120000 950000 1120000 n (Patient) 69 257 67 259 63 263 UO only Median 359000 582000 343000 586000 342000 567000 Average 431000 598000 413000 596000 411000 580000 Stdev 248000 287000 239000 285000 241000 283000 p (t-test) 5.3E−8 1.1E−9 1.9E−8 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 197 128 176 149 159 166 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.74 0.72 0.67 0.78 0.73 0.69 0.77 0.72 0.68 SE 0.032 0.031 0.031 0.032 0.031 0.030 0.034 0.032 0.030 p Value 7.8E−14 4.5E−13 8.1E−8 0 3.7E−14 5.0E−10 6.9E−15 1.3E−11 3.1E−9 nCohort Recovered 52 69 197 42 67 176 38 63 159 nCohort Non- 275 257 128 285 259 149 289 263 166 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 80% 82% 84% 80% 82% 85% 79% 81% 85% Specificity 52% 51% 31% 57% 52% 33% 58% 51% 35% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 56% 57% 62% 55% 57% 63% 54% 56% 60% Specificity 81% 75% 58% 83% 76% 61% 82% 75% 60% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 28% 29% 41% 28% 29% 40% 28% 29% 37% Specificity 92% 88% 85% 98% 90% 88% 97% 89% 87% OR Quartile 2 4.32 4.60 2.42 5.22 4.93 2.69 5.25 4.40 3.07 p Value 3.6E−6 1.4E−7 0.0021 1.7E−6 5.1E−8 3.6E−4 3.9E−6 6.1E−7 4.1E−5 Lower limit of 95% 2.33 2.61 1.38 2.65 2.78 1.56 2.60 2.46 1.79 CI Upper limit of 95% 8.02 8.12 4.26 10.3 8.76 4.64 10.6 7.87 5.24 CI OR Quartile 3 5.35 4.02 2.29 6.13 4.18 2.65 5.27 3.72 2.19 p Value 6.7E−6 5.6E−6 3.7E−4 2.6E−5 4.8E−6 2.2E−5 1.3E−4 3.0E−5 5.2E−4 Lower limit of 95% 2.58 2.21 1.45 2.64 2.27 1.69 2.25 2.01 1.41 CI Upper limit of 95% 11.1 7.34 3.61 14.3 7.72 4.16 12.4 6.90 3.42 CI OR Quartile 4 4.75 3.08 4.09 16.3 3.49 4.72 14.4 3.19 4.14 p Value 0.0037 0.0049 1.7E−7 0.0063 0.0030 4.0E−8 0.0090 0.0061 8.0E−7 Lower limit of 95% 1.66 1.41 2.41 2.20 1.53 2.71 1.94 1.39 2.36 CI Upper limit of 95% 13.6 6.76 6.94 120 7.99 8.21 107 7.32 7.29 CI

TABLE 6.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered″ and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 279000 525000 277000 499000 279000 490000 Average 363000 543000 366000 534000 370000 529000 Stdev 233000 275000 239000 275000 245000 275000 p (t-test) 1.0E−7 2.4E−6 1.7E−5 Min 66700 13600 66700 13600 66700 13600 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 87 240 76 251 69 258 sCr only Median 282000 528000 282000 528000 291000 521000 Average 380000 547000 376000 547000 382000 541000 Stdev 248000 273000 245000 273000 249000 274000 p (t-test) 3.0E−7 1.7E−7 1.8E−6 Min 66700 13600 66700 13600 66700 13600 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 101 226 99 228 94 233 UO only Median 371000 598000 369000 576000 371000 534000 Average 437000 617000 434000 595000 433000 578000 Stdev 250000 287000 248000 289000 245000 292000 p (t-test) 1.4E−8 1.9E−7 1.8E−6 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 217 108 198 127 181 144 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.69 0.68 0.69 0.69 0.66 0.68 0.68 0.65 SE 0.030 0.030 0.033 0.032 0.030 0.031 0.033 0.031 0.031 p Value 4.7E−11 4.8E−10 1.9E−8 4.3E−9 1.6E−10 2.4E−7 6.9E−8 7.1E−9 2.1E−6 nCohort Recovered 87 101 217 76 99 198 69 94 181 nCohort Non-recovered 240 226 108 251 228 127 258 233 144 Cutoff Quartile 2 260000 260000 262000 260000 262000 262000 260000 260000 262000 Sensitivity 81% 82% 87% 80% 82% 85% 80% 82% 84% Specificity 43% 42% 31% 43% 42% 31% 43% 41% 32% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 57% 58% 65% 56% 58% 62% 55% 57% 58% Specificity 70% 67% 57% 68% 68% 58% 68% 67% 56% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 30% 31% 44% 29% 31% 39% 29% 30% 37% Specificity 90% 87% 84% 89% 88% 84% 88% 87% 84% OR Quartile 2 3.21 3.31 3.00 3.16 3.46 2.59 3.05 3.13 2.48 p Value 1.9E−5 7.2E−6 6.5E−4 4.1E−5 3.5E−6 0.0011 1.1E−4 2.2E−5 0.0011 Lower limit of 95% CI 1.88 1.96 1.60 1.82 2.05 1.46 1.73 1.85 1.44 Upper limit of 95% CI 5.47 5.58 5.64 5.49 5.85 4.59 5.36 5.31 4.28 OR Quartile 3 3.17 2.84 2.46 2.73 2.88 2.23 2.62 2.70 1.81 p Value 1.7E−5 3.3E−5 2.3E−4 2.9E−4 3.0E−5 5.5E−4 8.1E−4 1.0E−4 0.0088 Lower limit of 95% CI 1.88 1.74 1.52 1.59 1.75 1.42 1.49 1.64 1.16 Upper limit of 95% CI 5.37 4.65 3.96 4.71 4.73 3.52 4.59 4.47 2.81 OR Quartile 4 3.79 2.98 4.01 3.55 3.21 3.37 3.07 2.93 3.05 p Value 4.4E−4 9.7E−4 2.2E−7 0.0015 5.9E−4 4.6E−6 0.0051 0.0016 2.8E−5 Lower limit of 95% CI 1.80 1.56 2.37 1.63 1.65 2.00 1.40 1.51 1.81 Upper limit of 95% CI 7.96 5.68 6.77 7.76 6.25 5.66 6.72 5.72 5.14

TABLE 6.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered'’ and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or scrum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 280000 560000 279000 530000 279000 524000 Average 362000 564000 364000 557000 366000 547000 Stdev 224000 276000 229000 275000 235000 275000 p (t-test) 1.2E−10 1.5E−9 4.4E−8 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 111 216 104 223 94 233 sCr only Median 286000 564000 282000 564000 286000 552000 Average 376000 568000 372000 568000 376000 561000 Stdev 238000 273000 236000 273000 239000 274000 p (t-test) 4.0E−10 2.0E−10 3.0E−9 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 124 203 121 206 116 211 UO only Median 367000 602000 363000 595000 367000 577000 Average 434000 627000 435000 608000 429000 596000 Stdev 247000 288000 249000 288000 243000 292000 p (t-test) 1.4E−9 2.8E−8 4.6E−8 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 219 106 208 117 193 132 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.71 0.70 0.71 0.71 0.68 0.70 0.70 0.67 SE 0.028 0.028 0.032 0.029 0.028 0.032 0.030 0.029 0.031 p Value 1.2E−14 1.7E−13 1.4E−9 2.1E−13 3.3E−14 2.6E−8 2.1E−11 1.9E−12 5.2E−8 nCohort Recovered 111 124 219 104 121 208 94 116 193 nCohort Non- 216 203 106 223 206 117 233 211 132 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 84% 84% 88% 83% 84% 86% 82% 84% 86% Specificity 42% 40% 31% 43% 41% 31% 44% 41% 32% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 60% 61% 66% 59% 61% 65% 58% 60% 62% Specificity 68% 67% 58% 69% 68% 58% 69% 67% 58% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 33% 33% 42% 32% 33% 40% 31% 33% 39% Specificity 91% 89% 83% 90% 89% 83% 89% 89% 84% OR Quartile 2 3.80 3.61 3.22 3.83 3.83 2.87 3.62 3.67 2.81 p Value 5.5E−7 1.4E−6 4.0E−4 5.1E−7 4.8E−7 6.2E−4 1.9E−6 9.7E−7 3.9E−4 Lower limit of 95% 2.25 2.14 1.69 2.27 2.27 1.57 2.13 2.18 1.59 CI Upper limit of 95% 6.40 6.08 6.15 6.48 6.46 5.25 6.15 6.19 4.99 CI OR Quartile 3 3.22 3.11 2.63 3.26 3.24 2.58 3.09 3.04 2.27 p Value 2.2E−6 2.0E−6 8.5E−5 2.8E−6 LIE−6 7.6E−5 1.4E−5 4.5E−6 4.0E−4 Lower limit of 95% 1.98 1.95 1.63 1.99 2.02 1.61 1.86 1.89 1.44 CI Upper limit of 95% 5.22 4.97 4.27 5.35 5.21 4.12 5.14 4.89 3.57 CI OR Quartile 4 5.05 3.96 3.63 4.48 4.18 3.32 3.76 3.85 3.53 p Value 7.5E−6 1.8E−5 1.3E−6 3.4E−5 1.3E−5 5.7E−6 2.7E−4 4.2E−5 2.3E−6 Lower limit of 95% 2.49 2.11 2.15 2.20 2.20 1.98 1.84 2.02 2.09 CI Upper limit of 95% 10.3 7.42 6.12 9.11 7.97 5.57 7.66 7.34 5.96 CI

TABLE 6.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or scrum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 296000 564000 293000 536000 279000 529000 Average 383000 573000 381000 566000 372000 561000 Stdev 232000 278000 233000 277000 236000 274000 p (t-test) 3.3E−10 1.7E−9 1.6E−9 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 134 193 125 202 113 214 sCr only Median 293000 567000 289000 567000 282000 565000 Average 381000 580000 377000 581000 376000 577000 Stdev 234000 275000 231000 275000 233000 274000 p (t-test) 3.6E−11 1.0E−11 2.7E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 139 188 137 190 133 194 UO only Median 373000 595000 371000 582000 369000 576000 Average 438000 621000 437000 609000 432000 599000 Stdev 249000 290000 248000 291000 246000 291000 p (t-test) 1.1E−8 4.2E−8 5.9E−8 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 220 105 211 114 198 127 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.71 0.69 0.70 0.72 0.68 0.71 0.72 0.67 SE 0.028 0.028 0.033 0.029 0.028 0.032 0.029 0.028 0.031 p Value 7.5E−13 2.6E−14 1.2E−8 2.5E−12 2.7E−15 5.0E−8 2.2E−13 6.7E−15 4.9E−8 nCohort Recovered 134 139 220 125 137 211 113 133 198 nCohort Non- 193 188 105 202 190 114 214 194 127 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 85% 86% 90% 85% 86% 88% 85% 86% 87% Specificity 40% 40% 32% 42% 40% 32% 44% 41% 33% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 60% 62% 65% 59% 62% 64% 59% 62% 62% Specificity 64% 65% 57% 65% 66% 57% 67% 67% 58% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 35% 36% 41% 34% 36% 40% 33% 35% 39% Specificity 90% 89% 82% 90% 90% 83% 89% 89% 84% OR Quartile 2 3.98 3.90 3.99 4.08 4.05 3.32 4.51 4.05 3.39 p Value 3.1E−7 4.9E−7 7.7E−5 1.6E−7 2.5E−7 1.9E−4 2.2E−8 2.2E−7 7.1E−5 Lower limit of 95% 2.34 2.30 2.01 2.41 2.38 1.77 2.66 2.39 1.86 CI Upper limit of 95% 6.75 6.64 7.92 6.91 6.89 6.24 7.65 6.88 6.19 CI OR Quartile 3 2.70 3.05 2.42 2.69 3.24 2.39 3.00 3.28 2.23 p Value 2.0E−5 1.7E−6 3.2E−4 2.6E−5 5.4E−7 2.7E−4 6.8E−6 5.0E−7 5.5E−4 Lower limit of 95% 1.71 1.93 1.49 1.70 2.05 1.50 1.86 2.06 1.42 CI Upper limit of 95% 4.26 4.82 3.91 4.28 5.14 3.83 4.84 5.21 3.52 CI OR Quartile 4 4.66 4.58 3.22 4.47 4.90 3.29 4.09 4.59 3.37 p Value 1.5E−6 1.2E−6 1.1E−5 5.1E−6 6.9E−7 6.8E−6 3.1E−5 1.9E−6 4.6E−6 Lower limit of 95% 2.49 2.48 1.91 2.35 2.61 1.96 2.11 2.45 2.00 CI Upper limit of 95% 8.73 8.45 5.42 8.51 9.17 5.52 7.94 8.59 5.66 CI

TABLE 6.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 359000 572000 342000 576000 342000 577000 Average 421000 595000 416000 597000 415000 598000 Stdev 240000 290000 237000 289000 237000 289000 p (t-test) 8.6E−9 1.6E−9 1.1E−9 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 187 140 184 143 183 144 sCr only Median 313000 564000 308000 564000 308000 564000 Average 402000 583000 399000 583000 399000 581000 Stdev 240000 280000 239000 279000 239000 279000 p (t-test) 1.3E−9 5.5E−10 8.4E−10 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 1159 168 156 171 154 173 UO only Median 375000 582000 369000 586000 359000 596000 Average 436000 616000 433000 617000 428000 622000 Stdev 245000 296000 243000 294000 241000 292000 p (t-test) 1.4E−8 4.2E−9 3.7E−10 Min 13600 135000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 215 110 212 113 209 116 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.68 0.69 0.68 0.68 0.70 0.69 0.69 0.69 0.70 SE 0.030 0.029 0.032 0.030 0.029 0.032 0.030 0.029 0.031 p Value 6.3E−9 3.9E−11 3.0E−8 8.4E−10 8.0E−12 6.3E−9 4.4E−10 1.5E−11 4.5E−10 nCohort Recovered 187 159 215 184 156 212 183 154 209 nCohort Non- 140 168 110 143 171 113 144 173 116 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 86% 86% 88% 87% 87% 88% 87% 86% 89% Specificity 34% 37% 32% 34% 38% 32% 34% 38% 33% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 62% 61% 65% 62% 62% 65% 62% 62% 66% Specificity 59% 62% 57% 59% 63% 58% 60% 63% 58% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 39% 37% 40% 40% 37% 41% 40% 36% 42% Specificity 86% 87% 82% 86% 88% 83% 86% 88% 84% OR Quartile 2 3.24 3.72 3.45 3.40 3.91 3.63 3.45 3.75 3.82 p Value 5.5E−5 2.3E−6 1.7E−4 2.7E−5 9.5E−7 9.1E−5 2.1E−5 1.6E−6 4.7E−5 Lower limit of 95% 1.83 2.16 1.81 1.92 2.27 1.90 1.95 2.18 2.00 CI Upper limit of 95% 5.73 6.41 6.59 6.01 6.75 6.93 6.11 6.44 7.29 CI OR Quartile 3 2.35 2.55 2.43 2.40 2.76 2.47 2.45 2.76 2.66 p Value 2.0E−4 4.0E−5 2.4E−4 1.3E−4 9.3E−6 1.7E−4 8.6E−5 9.2E−6 4.6E−5 Lower limit of 95% 1.50 1.63 1.51 1.53 1.76 1.54 1.57 1.76 1.66 CI Upper limit of 95% 3.67 3.98 3.91 3.75 4.31 3.97 3.84 4.32 4.27 CI OR Quartile 4 3.83 4.07 3.11 4.22 4.21 3.36 4.14 4.07 3.90 p Value 6.8E−7 1.1E−6 1.8E−5 1.6E−7 8.4E−7 4.8E−6 2.3E−7 1.5E−6 3.5E−7 Lower limit of 95% 2.26 2.31 1.85 2.46 2.37 2.00 2.42 2.30 2.31 CI Upper limit of 95% 6.52 7.14 5.21 7.22 7.45 5.64 7.09 7.21 6.58 CI

Example 7. Use of Metalloproteinase Inhibitor 1 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 1 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 7.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 7070 20000 6950 20000 6710 20500 Average 15900 35500 14900 35600 14800 35300 Stdev 23300 35800 21500 35900 21900 35700 p (t-test) 4.7E−7 1.2E−7 2.2E−7 Min 475 297 475 297 475 297 Max 116000 146000 110000 146000 110000 146000 n (Patient) 108 209 104 213 100 217 sCr only Median 8650 20500 8510 20500 8310 20500 Average 16400 36800 16400 36600 16500 36400 Stdev 22300 36800 22400 36700 22600 36500 p (t-test) 5.2E−8 8.5E−8 1.3E−7 Min 475 297 475 297 475 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 126 190 124 192 122 194 UO only Median 11300 21700 11000 21300 11000 20500 Average 24700 39200 24000 38700 24000 37300 Stdev 31100 36500 30700 36400 31000 35800 P (t-test) 3.9E−4 1.9E−4 6.3E−4 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n (Patient) 222 93 208 107 198 117 Recovery Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.72 0.70 0.66 0.73 0.70 0.66 0.73 0.70 0.65 SE 0.029 0.029 0.035 0.028 0.029 0.033 0.028 0.029 0.033 p Value 3.8E−14 9.9E−12 6.3E−6 2.2E−15 8.2E−12 1.7E−6 2.2E−16 6.5E−12 4.3E−6 nCohort Recovered 108 126 222 104 124 208 100 122 198 nCohort Non-recovered 209 190 93 213 192 107 217 194 117 Cutoff Quartile 2 6030 6020 6080 6030 6020 6080 6030 6020 6080 Sensitivity 85% 84% 89% 85% 84% 89% 85% 85% 87% Specificity 44% 39% 31% 45% 40% 32% 46% 40% 32% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity 63% 64% 68% 62% 64% 66% 63% 63% 64% Specificity 74% 71% 57% 75% 71% 58% 77% 71% 58% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity 33% 35% 35% 33% 34% 36% 32% 34% 35% Specificity 89% 90% 79% 90% 90% 81% 90% 89% 81% OR Quartile 2 4.59 3.39 3.74 4.66 3.53 3.76 4.75 3.67 3.25 p Value 2.8E−8 6.1E−6 3.0E−4 2.1E−8 3.2E−6 9.9E−5 1.6E−8 1.6E−6 1.9E−4 Lower limit of 95% CI 2.68 2.00 1.83 2.72 2.08 1.93 2.77 2.16 1.75 Upper limit of 95% CI 7.87 5.76 7.65 7.99 6.00 7.33 8.15 6.24 6.03 OR Quartile 3 4.80 4.22 2.81 4.99 4.26 2.74 5.62 4.31 2.47 p Value 2.2E−9 5.6E−9 7.2E−5 1.8E−9 5.3E−9 4.8E−5 3.9E−10 4.9E−9 1.7E−4 Lower limit of 95% CI 2.87 2.60 1.69 2.96 2.62 1.69 3.27 2.64 1.54 Upper limit of 95% CI 8.02 6.85 4.67 8.42 6.93 4.46 9.65 7.02 3.96 OR Quartile 4 3.86 4.63 2.10 4.60 4.47 2.41 4.29 4.32 2.27 p Value 7.1E−5 3.5E−6 0.0064 2.6E−5 5.7E−6 9.9E−4 6.3E−5 9.3E−6 0.0019 Lower limit of 95% CI 1.98 2.42 1.23 2.26 2.34 1.43 2.10 2.26 1.35 Upper limit of 95% CI 7.51 8.84 3.59 9.38 8.54 4.07 8.74 8.26 3.82

TABLE 7.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9010 20500 8810 20500 8420 20500 Average 17700 37200 17600 37000 16900 37000 Stdev 23400 37100 23500 37000 22300 37100 p (t-test) 1.6E−7 1.9E−7 6.6E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 110000 146000 n (Patient) 136 181 134 183 129 188 sCr only Median 9350 20500 9380 20500 9380 20500 Average 18000 38400 18100 38200 18100 38000 Stdev 22800 38000 22900 38000 23000 37800 p (t-test) 2.8E−8 4.1E−8 6.8E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 149 168 148 169 146 171 UO only Median 11900 21700 11800 21300 11200 20900 Average 25200 38600 25300 37400 24900 36700 Stdev 31100 37200 31400 36500 31200 36200 p (t-test) 0.0012 0.0030 0.0029 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n (Patient) 226 89 220 95 207 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.68 0.64 0.69 0.68 0.64 0.70 0.68 0.64 SE 0.029 0.030 0.036 0.029 0.030 0.035 0.029 0.030 0.034 p Value  9.9E−11 1.0E−9 7.8E−5  5.0E−11 1.4E−9 9.6E−5  2.3E−12 1.4E−9 5.6E−5 nCohort Recovered 136 149 226 134 148 220 129 146 207 nCohort Non- 181 168 89 183 169 95 188 171 108 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 86% 86% 90% 86% 86% 89% 86% 86% 88% Specificity 40% 37% 31% 40% 37% 31% 41% 38% 32% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 63% 64% 66% 63% 63% 65% 63% 63% 64% Specificity 67% 65% 56% 67% 65% 56% 69% 65% 57% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 34% 36% 33% 34% 36% 33% 34% 36% 32% Specificity 87% 87% 78% 87% 87% 78% 88% 87% 79% OR Quartile 2 4.11 3.51 3.99 4.27 3.57 3.88 4.35 3.70 3.42 p Value 3.6E−7 6.4E−6 2.7E−4 1.8E−7 4.8E−6 2.0E−4 1.1E−7 2.6E−6 2.0E−4 Lower limit 2.38 2.03 1.89 2.47 2.07 1.90 2.53 2.14 1.79 of 95% CI Upper limit 7.08 6.06 8.40 7.36 6.17 7.94 7.48 6.39 6.55 of 95% CI OR Quartile 3 3.44 3.27 2.52 3.46 3.19 2.43 3.84 3.19 2.35 p Value 2.2E−7 4.6E−7 4.0E−4 2.1E−7 8.0E−7 5.0E−4 3.2E−8 7.9E−7 4.9E−4 Lower limit 2.16 2.06 1.51 2.16 2.01 1.47 2.38 2.01 1.45 of 95% CI Upper limit 5.49 5.19 4.21 5.53 5.05 4.00 6.18 5.06 3.79 of 95% CI OR Quartile 4 3.42 3.90 1.70 3.61 3.83 1.74 3.65 3.71 1.78 p Value 3.7E−5 3.5E−6 0.055 2.2E−5 4.6E−6 0.043 2.7E−5 7.9E−6 0.031 Lower limit 1.91 2.19 0.988 2.00 2.16 1.02 1.99 2.09 1.05 of 95% CI Upper limit 6.12 6.93 2.93 6.54 6.81 2.96 6.67 6.59 3.00 of 95% CI

TABLE 7.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9010 21900 9010 21700 8870 21700 Average 18400 38700 18500 38300 18000 38300 Stdev 24700 37300 24900 37200 23900 37400 p (t-test) 3.1E−8 7.1E−8 3.0E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 111000 146000 n (Patient) 154 163 152 165 148 169 sCr only Median 9310 22000 9160 21900 9310 21600 Average 18700 39200 18800 39000 18900 38800 Stdev 24500 37800 24600 37800 24700 37700 p (t-test) 2.3E−8 3.4E−8 5.2E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 161 156 160 157 159 158 UO only Median 11900 21300 11900 20900 11800 20500 Average 25600 37100 25700 36700 25300 36500 Stdev 31500 36700 31500 36600 31200 36500 p (t-test) 0.0056 0.0077 0.0053 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n (Patient) 224 91 223 92 212 103 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.69 0.63 0.70 0.69 0.62 0.70 0.69 0.62 SE 0.029 0.030 0.036 0.029 0.030 0.036 0.029 0.030 0.034 p Value  4.1E−12  8.7E−11 3.1E−4  1.3E−11  1.3E−10 6.4E−4  3.0E−12  2.1E−10 3.7E−4 nCohort Recovered 154 161 224 152 160 223 148 159 212 nCohort Non- 163 156 91 165 157 92 169 158 103 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 87% 86% 88% 87% 86% 87% 86% 86% 86% Specificity 37% 35% 30% 38% 36% 30% 38% 36% 31% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 66% 66% 65% 65% 66% 64% 65% 65% 63% Specificity 67% 65% 56% 66% 65% 56% 67% 65% 56% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 37% 38% 32% 36% 38% 32% 36% 37% 32% Specificity 87% 87% 78% 87% 87% 78% 87% 87% 78% OR Quartile 2 3.77 3.34 3.17 3.90 3.40 2.86 3.86 3.45 2.81 p Value 2.9E−6 2.0E−5 0.0011 1.6E−6 1.6E−5 0.0021 1.5E−6 1.2E−5 0.0014 Lower limit 2.16 1.92 1.59 2.24 1.95 1.46 2.23 1.98 1.49 of 95% CI Upper limit 6.57 5.81 6.33 6.80 5.91 5.60 6.70 6.02 5.30 of 95% CI OR Quartile 3 3.97 3.64 2.33 3.75 3.54 2.24 3.77 3.44 2.19 p Value 7.3E−9 4.6E−8 0.0010 2.5E−8 8.4E−8 0.0016 2.4E−8 1.5E−7 0.0015 Lower limit 2.49 2.29 1.41 2.36 2.23 1.36 2.36 2.17 1.35 of 95% CI Upper limit 6.33 5.79 3.86 5.97 5.63 3.70 6.00 5.46 3.55 of 95% CI OR Quartile 4 3.90 4.05 1.63 3.77 3.98 1.59 3.83 3.92 1.70 p Value 2.6E−6 1.0E−6 0.078 4.5E−6 1.4E−6 0.092 4.6E−6 1.8E−6 0.048 Lower limit 2.21 2.31 0.947 2.14 2.27 0.927 2.16 2.23 1.00 of 95% CI Upper limit 6.88 7.11 2.80 6.65 6.98 2.74 6.81 6.86 2.88 of 95% CI

TABLE 7.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9350 21900 9360 21500 9310 21500 Average 19200 39700 19400 39100 18900 39100 Stdev 24900 38100 25100 37900 24200 38100 p (t-test) 2.3E−8 7.9E−7 3.5E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 111000 146000 n (Patient) 168 149 165 152 161 156 sCr only Median 9510 22000 9570 22000 9620 21900 Average 19400 40000 19500 39700 19600 39500 Stdev 24900 38400 24900 38400 25000 38300 p (t-test) 2.4E−8 3.7E−8 5.7E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 172 145 171 146 170 147 UO only Median 11800 21500 11900 21300 11800 20500 Average 25400 37400 25500 37200 25200 37000 Stdev 31500 36500 31500 36400 31100 36700 p (t-test) 0.0036 0.0046 0.0033 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n (Patient) 223 92 222 93 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.69 0.63 0.69 0.69 0.63 0.69 0.68 0.63 SE 0.030 0.030 0.036 0.030 0.030 0.035 0.029 0.030 0.034 p Value  3.9E−11  3.8E−10 1.9E−4  1.5E−10  5.6E−10 2.3E−4  4.7E−11  9.1E−10 1.6E−4 nCohort Recovered 168 172 223 165 171 222 161 170 214 nCohort Non- 149 145 92 152 146 93 156 147 101 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 87% 86% 88% 88% 86% 88% 87% 86% 88% Specificity 36% 34% 30% 36% 35% 31% 37% 35% 31% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 68% 68% 65% 66% 67% 65% 66% 67% 63% Specificity 65% 65% 56% 65% 64% 56% 65% 64% 56% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 38% 38% 35% 37% 38% 34% 37% 37% 34% Specificity 86% 85% 79% 85% 85% 79% 86% 85% 79% OR Quartile 2 3.80 3.26 3.23 4.00 3.32 3.29 3.93 3.38 3.38 p Value 5.4E−6 4.4E−5 8.9E−4 2.4E−6 3.4E−5 7.3E−4 2.3E−6 2.7E−5 3.5E−4 Lower limit 2.14 1.85 1.62 2.25 1.88 1.65 2.23 1.91 1.73 of 95% CI Upper limit 6.76 5.76 6.45 7.12 5.85 6.57 6.94 5.95 6.60 of 95% CI OR Quartile 3 3.99 3.79 2.39 3.65 3.68 2.30 3.64 3.57 2.21 p Value 6.9E−9 2.3E−8 7.0E−4 4.5E−8 4.2E−8 0.0011 4.6E−8 7.7E−8 0.0014 Lower limit 2.50 2.38 1.44 2.30 2.31 1.39 2.29 2.25 1.36 of 95% CI Upper limit 6.37 6.06 3.96 5.81 5.87 3.80 5.79 5.69 3.59 of 95% CI OR Quartile 4 3.61 3.59 2.00 3.43 3.53 1.95 3.45 3.47 1.91 p Value 3.8E−6 3.5E−6 0.011 9.1E−6 4.8E−6 0.014 9.5E−6 6.4E−6 0.017 Lower limit 2.10 2.09 1.17 1.99 2.06 1.14 2.00 2.02 1.12 of 95% CI Upper limit 6.23 6.17 3.41 5.90 6.06 3.34 5.98 5.95 3.23 of 95% CI

TABLE 7.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11100 21500 11100 21100 11000 20800 Average 23100 39100 23200 38600 23300 38300 Stdev 28900 38100 29000 38000 29000 37900 p (t-test) 3.3E−5 6.5E−5 8.6E−5 Min 297 399 297 399 297 399 Max 127000 146000 127000 146000 127000 146000 n (Patient) 203 114 201 116 200 117 sCr only Median 10900 22000 10900 22000 10900 22000 Average 20400 40600 20400 40600 20400 40600 Stdev 25100 39400 25100 39400 25100 39400 p (t-test) 5.4E−8 5.4E−8 5.4E−8 Min 419 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 185 132 185 132 185 132 UO only Median 11800 21500 11900 21300 11900 20800 Average 24500 38800 24600 38500 24700 37900 Stdev 29700 38800 29800 38700 29900 38500 p (t-test) 4.1E−4 5.5E−4 9.8E−4 Min 297 399 297 399 297 399 Max 127000 146000 127000 146000 127000 146000 n (Patient) 217 98 216 99 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.67 0.63 0.65 0.67 0.63 0.64 0.67 0.63 SE 0.033 0.031 0.035 0.033 0.031 0.035 0.033 0.031 0.035 p Value 4.4E−6 2.0E−8 1.3E−4 8.7E−6 2.0E−8 1.5E−4 9.7E−6 2.0E−8 2.5E−4 nCohort Recovered 203 185 217 201 185 216 200 185 214 nCohort Non- 114 132 98 116 132 99 117 132 101 recovered Cutoff Quartile 2 6030 6030 6080 6030 6030 6080 6030 6030 6080 Sensitivity 86% 87% 87% 86% 87% 87% 86% 87% 87% Specificity 31% 34% 30% 31% 34% 31% 32% 34% 31% Cutoff Quartile 3 14200 14200 14200 14200 14200 14200 14200 14200 14200 Sensitivity 66% 65% 64% 65% 65% 64% 64% 65% 62% Specificity 59% 61% 56% 58% 61% 56% 58% 61% 56% Cutoff Quartile 4 35900 35900 36300 35900 35900 36300 35900 35900 36300 Sensitivity 37% 39% 35% 36% 39% 34% 36% 39% 34% Specificity 81% 84% 79% 81% 84% 79% 81% 84% 79% OR Quartile 2 2.76 3.41 2.86 2.85 3.41 2.91 2.90 3.41 3.02 p Value 0.0010 5.1E−5 0.0016 7.0E−4 5.1E−5 0.0013 5.7E−4 5.1E−5 8.7E−4 Lower limit 1.50 1.88 1.49 1.56 1.88 1.52 1.58 1.88 1.57 of 95% CI Upper limit 5.05 6.17 5.48 5.23 6.17 5.58 5.32 6.17 5.79 of 95% CI OR Quartile 3 2.72 2.87 2.31 2.55 2.87 2.23 2.47 2.87 2.08 p Value 3.9E−5 8.5E−6 8.6E−4 1.1E−4 8.5E−6 0.0013 1.7E−4 8.5E−6 0.0031 Lower limit 1.69 1.80 1.41 1.59 1.80 1.37 1.54 1.80 1.28 of 95% CI Upper limit 4.39 4.56 3.78 4.09 4.56 3.64 3.95 4.56 3.37 of 95% CI OR Quartile 4 2.53 3.39 2.03 2.43 3.39 1.99 2.39 3.39 1.91 p Value 4.5E−4 6.2E−6 0.0088 7.6E−4 6.2E−6 0.011 9.7E−4 6.2E−6 0.017 Lower limit 1.51 2.00 1.20 1.45 2.00 1.17 1.42 2.00 1.12 of 95% CI Upper limit 4.26 5.75 3.45 4.09 5.75 3.37 4.00 5.75 3.23 of 95% CI

TABLE 7.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 274000 564000 275000 564000 270000 564000 Average 354000 562000 354000 560000 342000 561000 Stdev 231000 271000 231000 271000 222000 271000 p (t-test) 5.2E−11 1.3E−10 1.0E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 105 222 102 225 98 229 sCr only Median 279000 566000 279000 566000 275000 566000 Average 365000 569000 362000 569000 358000 570000 Stdev 235000 269000 235000 268000 232000 268000 p (t-test) 2.1E−11 1.4E−11 4.4E−12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 121 205 119 207 117 209 UO only Median 373000 605000 371000 598000 369000 595000 Average 445000 617000 439000 608000 433000 603000 Stdev 256000 284000 252000 287000 249000 286000 p (t-test) 1.4E−7  7.4E−8  3.9E−8  Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 226 99 213 112 202 123 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.72 0.67 0.73 0.73 0.67 0.74 0.73 0.67 SE 0.028 0.028 0.034 0.028 0.028 0.032 0.028 0.027 0.031 p Value  2.2E−16  4.4E−16 3.0E−7  1.3E−15 0 1.3E−7 0 0 4.0E−8 nCohort Recovered 105 121 226 102 119 213 98 117 202 nCohort Non- 222 205 99 225 207 112 229 209 123 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 84% 85% 85% 84% 86% 85% 84% 86% 85% Specificity 45% 43% 29% 44% 44% 30% 46% 44% 31% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 60% 62% 66% 60% 62% 64% 60% 62% 63% Specificity 71% 70% 57% 72% 71% 57% 72% 71% 57% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 33% 34% 44% 32% 33% 42% 33% 33% 41% Specificity 91% 89% 83% 91% 89% 84% 93% 90% 84% OR Quartile 2 4.33 4.40 2.31 4.01 4.58 2.40 4.41 4.77 2.64 p Value 5.2E−8 4.0E−8 0.0081 2.3E−7 1.8E−8 0.0038 4.3E−8 8.2E−9 0.0011 Lower limit 2.55 2.59 1.24 2.37 2.70 1.33 2.59 2.81 1.48 of 95% CI Upper limit 7.34 7.46 4.29 6.79 7.78 4.35 7.49 8.12 4.73 of 95% CI OR Quartile 3 3.81 3.84 2.50 3.78 3.89 2.41 3.92 3.94 2.26 p Value 1.8E−7 4.1E−8 2.6E−4 2.7E−7 3.8E−8 2.6E−4 2.2E−7 3.5E−8 5.1E−4 Lower limit 2.31 2.38 1.53 2.28 2.40 1.50 2.34 2.42 1.43 of 95% CI Upper limit 6.29 6.22 4.08 6.26 6.31 3.87 6.56 6.41 3.57 of 95% CI OR Quartile 4 5.23 4.21 3.96 4.96 4.08 3.68 6.33 4.41 3.64 p Value 1.1E−5 1.2E−5 3.3E−7 2.1E−5 1.9E−5 1.0E−6 9.5E−6 1.2E−5 1.2E−6 Lower limit 2.50 2.21 2.33 2.37 2.14 2.18 2.80 2.27 2.16 of 95% CI Upper limit 10.9 8.03 6.71 10.4 7.77 6.20 14.3 8.55 6.13 of 95% CI

TABLE 7.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 293000 568000 293000 571000 284000 568000 Average 379000 576000 376000 577000 370000 576000 Stdev 235000 274000 234000 273000 229000 274000 p (t-test) 5.1E−11 2.7E−11 8.5E−12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 134 193 133 194 128 199 sCr only Median 300000 576000 299000 577000 294000 577000 Average 386000 583000 384000 584000 380000 585000 Stdev 237000 274000 237000 274000 235000 273000 p (t-test) 3.5E−11 1.9E−11 6.9E−12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 146 181 145 182 143 184 UO only Median 375000 598000 375000 598000 369000 596000 Average 448000 619000 443000 618000 432000 616000 Stdev 256000 287000 253000 288000 247000 288000 p (t-test) 2.5E−7  7.7E−8  4.4E−9  Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 231 94 225 100 210 115 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.71 0.67 0.72 0.72 0.68 0.72 0.72 0.69 SE 0.028 0.028 0.034 0.028 0.028 0.033 0.028 0.028 0.032 p Value  1.9E−14  4.0E−14 3.9E−7  5.3E−15  1.2E−14 9.9E−8  6.7E−16  2.0E−15 4.2E−9 nCohort Recovered 134 146 231 133 145 225 128 143 210 nCohort Non- 193 181 94 194 182 100 199 184 115 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 85% 86% 87% 86% 86% 88% 85% 86% 88% Specificity 40% 39% 30% 41% 39% 31% 41% 40% 32% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 62% 63% 65% 62% 63% 64% 62% 63% 63% Specificity 67% 66% 56% 68% 66% 56% 68% 66% 57% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 35% 36% 43% 35% 36% 42% 35% 36% 42% Specificity 89% 88% 82% 89% 88% 82% 90% 89% 84% OR Quartile 2 3.98 4.00 2.91 4.05 4.07 3.24 4.14 4.22 3.38 p Value 3.1E−7 4.4E−7 0.0017 2.2E−7 3.2E−7 5.4E−4 1.3E−7 1.6E−7 1.5E−4 Lower limit 2.34 2.33 1.49 2.39 2.38 1.67 2.44 2.46 1.80 of 95% CI Upper limit 6.75 6.84 5.68 6.88 6.96 6.32 7.02 7.22 6.35 of 95% CI OR Quartile 3 3.36 3.27 2.34 3.47 3.36 2.26 3.43 3.38 2.32 p Value 2.9E−7 3.6E−7 8.1E−4 1.6E−7 2.0E−7 9.9E−4 2.5E−7 2.0E−7 4.3E−4 Lower limit 2.12 2.07 1.42 2.18 2.13 1.39 2.15 2.13 1.45 of 95% CI Upper limit 5.34 5.15 3.84 5.52 5.31 3.68 5.49 5.34 3.70 of 95% CI OR Quartile 4 4.22 4.25 3.33 4.15 4.18 3.35 4.70 4.44 3.71 p Value 4.2E−6 1.5E−6 7.9E−6 5.4E−6 2.0E−6 6.1E−6 2.5E−6 1.2E−6 8.5E−7 Lower limit 2.28 2.36 1.97 2.25 2.32 1.98 2.47 2.43 2.20 of 95% CI Upper limit 7.79 7.67 5.65 7.66 7.54 5.66 8.94 8.10 6.25 of 95% CI

TABLE 7.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 303000 573000 300000 576000 296000 576000 Average 394000 586000 390000 587000 384000 587000 Stdev 239000 276000 238000 275000 232000 276000 p (t-test) 1.0E−10 2.8E−11 7.0E−12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 154 173 152 175 148 179 sCr only Median 304000 582000 303000 586000 301000 590000 Average 394000 594000 392000 594000 389000 596000 Stdev 239000 275000 238000 274000 236000 274000 p (t-test) 1.4E−11 7.8E−12 2.2E−12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 161 166 160 167 159 168 UO only Median 373000 600000 373000 600000 367000 597000 Average 443000 627000 442000 627000 434000 621000 Stdev 254000 285000 252000 287000 248000 287000 p (t-test) 2.5E−8  1.7E−8  3.3E−9  Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 230 95 228 97 215 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.71 0.69 0.71 0.72 0.69 0.72 0.72 0.69 SE 0.028 0.028 0.034 0.028 0.028 0.034 0.028 0.028 0.032 p Value  3.2E−13  3.4E−14 2.8E−8  3.4E−14  1.1E−14 2.3E−8  4.9E−15  2.0E−15 2.6E−9 nCohort Recovered 154 161 230 152 160 228 148 159 215 nCohort Non- 173 166 95 175 167 97 179 168 110 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 87% 88% 88% 87% 88% 88% 87% 88% 88% Specificity 39% 39% 30% 39% 39% 30% 40% 39% 32% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 62% 63% 66% 62% 63% 66% 62% 64% 65% Specificity 63% 63% 57% 64% 64% 57% 64% 64% 57% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 36% 37% 42% 36% 37% 42% 36% 37% 42% Specificity 87% 87% 82% 88% 87% 82% 89% 87% 83% OR Quartile 2 4.38 4.57 3.34 4.54 4.65 3.07 4.50 4.73 3.45 p Value 1.6E−7 1.3E−7 5.9E−4 8.3E−8 9.8E−8 9.7E−4 7.4E−8 7.2E−8 1.7E−4 Lower limit 2.52 2.60 1.68 2.61 2.64 1.58 2.60 2.69 1.81 of 95% CI Upper limit 7.61 8.04 6.65 7.88 8.18 5.99 7.78 8.32 6.59 of 95% CI OR Quartile 3 2.76 2.98 2.56 2.91 3.06 2.53 2.93 3.14 2.43 p Value 9.2E−6 2.0E−6 2.2E−4 3.3E−6 1.2E−6 2.4E−4 3.2E−6 6.9E−7 2.4E−4 Lower limit 1.76 1.90 1.55 1.86 1.95 1.54 1.86 2.00 1.51 of 95% CI Upper limit 4.32 4.67 4.22 4.57 4.80 4.15 4.60 4.93 3.91 of 95% CI OR Quartile 4 3.74 3.87 3.26 3.94 3.81 3.34 4.39 4.07 3.57 p Value 4.4E−6 1.9E−6 1.1E−5 2.6E−6 2.5E−6 7.0E−6 8.8E−7 1.1E−6 1.7E−6 Lower limit 2.13 2.22 1.92 2.22 2.18 1.97 2.44 2.31 2.12 of 95% CI Upper limit 6.57 6.76 5.51 6.97 6.65 5.65 7.93 7.14 6.02 of 95% CI

TABLE 7.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 318000 576000 313000 577000 304000 577000 Average 401000 592000 398000 592000 393000 593000 Stdev 243000 275000 243000 273000 238000 275000 p (t-test) 1.3E−10 4.8E−11 1.3E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 166 161 163 164 159 168 sCr only Median 313000 590000 311000 594000 309000 594000 Average 397000 600000 395000 601000 393000 603000 Stdev 242000 272000 242000 271000 240000 271000 p (t-test) 6.4E−12 3.5E−12 9.9E−13 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 169 158 168 159 167 160 UO only Median 377000 598000 375000 596000 371000 596000 Average 441000 626000 441000 624000 436000 621000 Stdev 251000 289000 251000 288000 249000 287000 p (t-test) 1.2E−8  1.7E−8  5.7E−9  Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 226 99 225 100 217 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.72 0.69 0.71 0.72 0.69 0.71 0.73 0.69 SE 0.029 0.028 0.033 0.028 0.028 0.033 0.028 0.028 0.032 p Value  8.9E−13  1.4E−14 1.3E−8  1.2E−13  4.9E−15 1.6E−8  2.3E−14  8.9E−16 3.9E−9 nCohort Recovered 166 169 226 163 168 225 159 167 217 nCohort Non- 161 158 99 164 159 100 168 160 108 recovered Cutoff Quartile 2 260000 260000 2600002 260000 260000 262000 260000 260000 262000 Sensitivity 88% 89% 90% 88% 89% 90% 88% 89% 90% Specificity 38% 38% 31% 39% 38% 32% 39% 38% 32% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 63% 65% 66% 63% 65% 65% 63% 66% 64% Specificity 63% 64% 57% 63% 64% 56% 64% 65% 57% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 37% 37% 41% 37% 37% 41% 37% 38% 42% Specificity 86% 86% 82% 87% 86% 82% 87% 87% 83% OR Quartile 2 4.57 4.74 4.08 4.81 4.82 4.15 4.73 4.90 4.20 p Value 1.9E−7 1.5E−7 1.1E−4 7.7E−8 1.1E−7 8.8E−5 7.2E−8 8.3E−8 4.1E−5 Lower limit 2.58 2.65 2.00 2.71 2.70 2.04 2.69 2.74 2.12 of 95% CI Upper limit 8.10 8.47 8.30 8.53 8.62 8.45 8.32 8.77 8.33 of 95% CI OR Quartile 3 2.90 3.32 2.50 2.98 3.40 2.41 2.98 3.49 2.32 p Value 3.4E−6 2.2E−7 2.6E−4 2.0E−6 1.3E−7 4.2E−4 2.0E−6 7.1E−8 5.4E−4 Lower limit 1.85 2.11 1.53 1.90 2.16 1.48 1.90 2.22 1.44 of 95% CI Upper limit 4.54 5.22 4.08 4.67 5.36 3.92 4.67 5.51 3.73 of 95% CI OR Quartile 4 3.60 3.78 3.19 3.70 3.72 3.12 4.07 3.95 3.47 p Value 4.1E−6 1.7E−6 1.4E−5 3.2E−6 2.3E−6 2.0E−5 1.1E−6 1.0E−6 2.8E−6 Lower limit 2.09 2.19 1.89 2.13 2.16 1.85 2.31 2.28 2.06 of 95% CI Upper limit 6.20 6.53 5.39 6.41 6.41 5.26 7.14 6.86 5.85 of 95% CI

TABLE 7.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 358000 578000 350000 578000 342000 582000 Average 428000 607000 426000 607000 424000 609000 Stdev 246000 288000 246000 286000 244000 286000 p (t-test) 5.2E−9 4.0E−9 1.4E−9 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 204 123 202 125 201 126 sCr only Median 328000 578000 328000 578000 328000 578000 Average 412000 603000 412000 603000 412000 603000 Stdev 242000 281000 242000 281000 242000 281000 p (t-test) 1.8E−10 1.8E−10 1.8E−10 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 184 143 184 143 184 143 UO only Median 377000 586000 375000 582000 371000 590000 Average 439000 619000 439000 617000 436000 620000 Stdev 247000 295000 247000 295000 246000 293000 p (t-test) 2.0E−8 2.8E−8 6.1E−9 Min 13600 135000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 220 105 219 106 217 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.68 0.70 0.68 0.69 0.70 0.68 0.69 0.70 0.69 SE 0.031 0.030 0.033 0.031 0.030 0.033 0.031 0.030 0.032 p Value 3.2E−9  1.9E−11 3.8E−8 1.8E−9  1.9E−11 4.5E−8  5.7E−10  1.9E−11 8.2E−9 nCohort Recovered 204 184 220 202 184 219 201 184 217 nCohort Non- 123 143 105 125 143 106 126 143 108 recovered Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 89% 88% 89% 89% 88% 89% 89% 88% 89% Specificity 33% 35% 31% 34% 35% 32% 34% 35% 32% Cutoff Quartile 3 437000 437000 437000 437000 437000 437000 437000 437000 437000 Sensitivity 64% 64% 65% 64% 64% 64% 64% 64% 65% Specificity 58% 61% 57% 58% 61% 57% 59% 61% 57% Cutoff Quartile 4 707000 707000 708000 707000 707000 708000 707000 707000 708000 Sensitivity 39% 38% 40% 39% 38% 40% 40% 38% 41% Specificity 83% 85% 82% 84% 85% 82% 84% 85% 82% OR Quartile 2 3.89 4.05 3.54 4.02 4.05 3.60 4.09 4.05 3.73 p Value 2.2E−5 3.4E−6 1.9E−4 1.4E−5 3.4E−6 1.6E−4 1.1E−5 3.4E−6 1.0E−4 Lower limit of 2.08 2.24 1.82 2.15 2.24 1.85 2.18 2.24 1.92 95% CI Upper limit of 7.29 7.30 6.89 7.54 7.30 7.01 7.66 7.30 7.25 95% CI OR Quartile 3 2.51 2.81 2.42 2.50 2.81 2.34 2.56 2.81 2.46 p Value 9.2E−5 7.8E−6 3.2E−4 9.7E−5 7.8E−6 5.1E−4 6.2E−5 7.8E−6 2.3E−4 Lower limit of 1.58 1.78 1.49 1.58 1.78 1.45 1.62 1.78 1.52 95% CI Upper limit of 3.99 4.41 3.91 3.96 4.41 3.77 4.05 4.41 3.96 95% CI OR Quartile 4 3.20 3.63 3.00 3.30 3.63 2.94 3.47 3.63 3.24 p Value 1.0E−5 1.8E−6 3.4E−5 6.1E−6 1.8E−6 4.7E−5 2.6E−6 1.8E−6 9.3E−6 Lower limit of 1.91 2.14 1.78 1.97 2.14 1.75 2.07 2.14 1.93 95% CI Upper limit of 5.36 6.17 5.04 5.54 6.17 4.93 5.84 6.17 5.44 95% CI

Example 8. Use of Metalloproteinase Inhibitor 1 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 1 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 8.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 10700 28600 10700 33800 11000 31700 Average 19900 45200 20100 48800 22000 48700 Stdev 25000 40000 25500 40200 27800 39800 p (t-test) 2.2E−11 2.4E−13  1.7E−10 Min 297 873 297 873 297 873 Max 127000 146000 127000 146000 127000 146000 n (Patient) 205 112 221 96 236 81 sCr only Median 11000 28600 10900 31500 11200 31500 Average 20000 45400 20300 48800 22200 47500 Stdev 24700 40600 24900 41400 27400 41100 p (t-test) 2.8E−11 6.1E−13 1.2E−9 Min 297 873 297 873 297 873 Max 127000 146000 127000 146000 127000 146000 n (Patient) 208 108 223 93 235 81 UO only Median 12400 25100 12500 28600 12700 30000 Average 26300 47200 26300 48500 26300 49300 Stdev 31200 42100 31100 43100 31000 43400 p (t-test) 2.0E−4  1.3E−4  8.7E−5 Min 297 873 297 873 297 873 Max 146000 140000 146000 140000 146000 140000 n (Patient) 275 40 278 37 279 36 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.70 0.67 0.74 0.73 0.67 0.74 0.71 0.67 SE 0.031 0.032 0.049 0.032 0.033 0.051 0.035 0.036 0.052 p Value  9.6E−12  2.8E−10 5.0E−4  2.0E−14  3.4E−12 7.3E−4  1.0E−11 5.7E−9 7.4E−4 nCohort Non- 205 208 275 221 223 278 236 235 279 persistent nCohort Persistent 112 108 40 96 93 37 81 81 36 Cutoff Quartile 2 6030 6020 6080 6030 6020 6080 6030 6020 6080 Sensitivity 89% 88% 92% 92% 90% 92% 93% 90% 92% Specificity 33% 32% 28% 32% 31% 27% 31% 30% 27% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity 71% 69% 70% 76% 74% 70% 78% 74% 69% Specificity 61% 60% 53% 61% 60% 53% 59% 58% 52% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity 46% 45% 45% 50% 48% 46% 49% 47% 47% Specificity 86% 86% 78% 86% 85% 78% 83% 83% 78% OR Quartile 2 4.05 3.40 4.71 5.21 4.27 4.26 5.60 3.95 4.12 p Value 3.9E−5 2.2E−4 0.012 3.1E−5 1.3E−4 0.019 1.2E−4 5.7E−4 0.022 Lower limit of 2.08 1.77 1.41 2.39 2.03 1.27 2.33 1.81 1.23 95% CI Upper limit of 7.88 6.50 15.7 11.3 8.98 14.3 13.4 8.63 13.8 95% CI OR Quartile 3 3.74 3.42 2.60 4.98 4.33 2.61 5.10 3.99 2.49 p Value 1.6E−7 1.1E−6 0.0089 6.0E−9 8.5E−8 0.011 4.6E−8 1.3E−6 0.016 Lower limit of 2.28 2.09 1.27 2.90 2.53 1.24 2.84 2.28 1.18 95% CI Upper limit of 6.13 5.61 5.33 8.56 7.40 5.50 9.16 7.00 5.27 95% CI OR Quartile 4 5.07 4.93 2.87 5.91 5.21 2.96 4.78 4.18 3.13 p Value 4.0E−9 7.8E−9 0.0025  2.1E−10 3.2E−9 0.0026 2.9E−8 3.7E−7 0.0017 Lower limit of 2.95 2.87 1.45 3.41 3.02 1.46 2.75 2.41 1.54 95% CI Upper limit of 8.72 8.47 5.69 10.2 9.00 6.00 8.31 7.26 6.39 95% CI

TABLE 8.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 10100 23400 10600 31500 10800 31500 Average 18900 42900 18900 46800 20700 46900 Stdev 23700 39500 23400 40500 26300 39900 p (t-test) 7.7E−11 9.3E−14  2.4E−11 Min 419 297 297 399 297 399 Max 121000 146000 121000 146000 121000 146000 n (Patient) 186 131 204 113 219 98 sCr only Median 11000 22400 10900 29300 11000 27300 Average 19500 42800 19400 46000 21400 44900 Stdev 23700 40400 23600 41200 26500 40600 p (t-test) 4.0E−10 2.3E−12 2.6E−9 Min 419 297 297 399 297 399 Max 121000 146000 121000 146000 121000 146000 n (Patient) 192 124 206 110 218 98 UO only Median 12400 23400 12400 23400 12400 24700 Average 26000 42000 26000 42700 26000 43100 Stdev 31400 38800 31200 39800 31200 40000 p (t-test) 8.4E−4  7.3E−4  5.6E−4 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n (Patient) 256 59 260 55 261 54 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.68 0.65 0.73 0.71 0.65 0.73 0.70 0.65 SE 0.030 0.031 0.042 0.031 0.032 0.043 0.032 0.033 0.044 p Value  2.2E−11 7.7E−9 3.0E−4  4.2E−14  2.8E−11 6.8E−4  1.0E−12 3.7E−9 7.2E−4 nCohort Non- 186 192 256 204 206 260 219 218 261 persistent nCohort Persistent 131 124 59 113 110 55 98 98 54 Cutoff Quartile 2 6030 6020 6080 6030 6020 6080 6030 6020 6080 Sensitivity 88% 86% 93% 90% 89% 93% 93% 91% 93% Specificity 34% 32% 29% 33% 33% 29% 33% 32% 29% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity 68% 66% 68% 73% 70% 67% 74% 70% 67% Specificity 62% 60% 54% 62% 61% 53% 61% 59% 53% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity 43% 42% 39% 48% 45% 40% 48% 44% 41% Specificity 87% 86% 78% 87% 86% 78% 85% 83% 78% OR Quartile 2 3.68 3.00 5.70 4.64 3.94 5.17 6.37 4.68 5.04 p Value 2.4E−5 2.9E−4 0.0012 1.2E−5 5.6E−5 0.0022 9.4E−6 4.6E−5 0.0026 Lower limit of 2.01 1.66 1.99 2.33 2.02 1.80 2.81 2.23 1.76 95% CI Upper limit of 6.74 5.44 16.3 9.21 7.67 14.8 14.4 9.82 14.4 95% CI OR Quartile 3 3.51 2.98 2.46 4.36 3.60 2.36 4.52 3.45 2.28 p Value 1.8E−7 5.6E−6 0.0032 8.2E−9 3.8E−7 0.0061 2.3E−8 2.0E−6 0.0088 Lower limit of 2.19 1.86 1.35 2.64 2.20 1.28 2.66 2.07 1.23 95% CI Upper limit of 5.63 4.77 4.48 7.20 5.90 4.36 7.66 5.75 4.22 95% CI OR Quartile 4 5.04 4.41 2.28 6.27 5.09 2.37 5.19 3.95 2.46 p Value 8.7E−9 7.5E−8 0.0072  7.7E−11 4.4E−9 0.0058 2.6E−9 5.0E−7 0.0042 Lower limit of 2.90 2.57 1.25 3.61 2.95 1.28 3.02 2.31 1.33 95% CI Upper limit of 8.74 7.58 4.16 10.9 8.76 4.39 8.93 6.75 4.56 95% CI

TABLE 8.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9450 22000 9580 28600 9980 27300 Average 18500 41600 18300 44600 20200 44700 Stdev 23100 39200 22800 39900 25200 40100 p (t-test) 2.5E−10 9.6E−13 1.2E−10 Min 419 297 297 399 297 399 Max 116000 146000 116000 146000 120000 146000 n 175 142 190 127 205 112 (Patient) sCr only Median 10600 22000 9980 28600 10800 22700 Average 18900 41900 18700 44700 20600 44200 Stdev 23000 40000 22800 40600 25200 40800 p (t-test) 4.4E−10 3.1E−12 8.4E−10 Min 419 297 297 399 297 399 Max 116000 146000 116000 146000 120000 146000 n 182 134 195 121 208 108 (Patient) UO only Median 12400 21900 12400 22000 12400 22000 Average 25800 40300 25800 41300 25800 41300 Stdev 31100 38700 30900 39700 30900 39700 p (t-test) 0.0014 8.6E−4 8.6E−4 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n 246 69 251 64 251 64 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.70   0.69   0.64   0.73   0.71   0.64   0.72   0.70   0.64 SE   0.030   0.031   0.039   0.030   0.031   0.041   0.031   0.032   0.041 p Value 1.2E−11 1.2E−9 5.0E−4 1.8E−14 3.1E−12 8.1E−4 6.4E−12 8.5E−10 8.1E−4 nCohort Non-  175  182  246  190  195  251  205  208  251 persistent nCohort Persistent  142  134   69  127  121   64  112  108   64 Cutoff Quartile 2  6030  6020  6080  6030  6020  6080  6030  6020  6080 Sensitivity   87%   86%   91%   90%   88%   91%   91%   90%   91% Specificity   35%   33%   30%   35%   33%   29%   34%   33%   29% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity   67%   66%   64%   70%   69%   64%   71%   69%   64% Specificity   63%   62%   54%   63%   62%   53%   61%   60%   53% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity   40%   40%   36%   44%   44%   38%   44%   43%   38% Specificity   87%   86%   78%   87%   87%   78%   85%   84%   78% OR Quartile 2   3.69   2.98   4.43   4.67   3.82   3.96   5.18   4.28   3.96 p Value 1.2E−5 2.0E−4 9.3E−4 3.0E−6 3.2E−5   0.0022 5.9E−6 3.4E−5   0.0022 Lower limit of 95% CI   2.06   1.67   1.84   2.44   2.03   1.64   2.54   2.15   1.64 Upper limit of 95% CI   6.61   5.29   10.7   8.91   7.19   9.59   10.5   8.52   9.59 OR Quartile 3   3.51   3.24   2.04   4.02   3.71   2.04   3.74   3.42   2.04 p Value 1.3E−7 8.2E−7   0.011 1.5E−8 1.0E−7   0.014 1.6E−7 1.1E−6   0.014 Lower limit of 95% CI   2.20   2.03   1.17   2.48   2.29   1.16   2.28   2.09   1.16 Upper limit of 95% CI   5.58   5.17   3.54   6.49   6.02   3.60   6.13   5.61   3.60 OR Quartile 4   4.43   4.24   2.02   5.46   5.07   2.14   4.37   3.93   2.14 p Value 1.3E−7 2.1E−7   0.017 1.8E−9 6.2E−9   0.011 6.4E−8 4.7E−7   0.011 Lower limit of 95% CI   2.55   2.46   1.14   3.14   2.93   1.19   2.56   2.31   1.19 Upper limit of 95% CI   7.70   7.31   3.59   9.48   8.76   3.85   7.45   6.70   3.85

TABLE 8.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9450 22000 9600 28600 9980 24700 Average 18300 41500 18400 44300 19800 44400 Stdev 22900 39100 22800 39900 24400 40500 p (t-test) 2.0E−10 2.1E−12 6.9E−11 Min 419 297 297 399 297 399 Max 116000 146000 116000 146000 116000 146000 n 173 144 189 128 201 116 (Patient) sCr only Median 10800 22000 10200 27300 10800 22700 Average 19100 41400 18800 43900 20200 44100 Stdev 23100 39900 22900 40400 24400 41100 p (t-test) 1.4E−9 1.3E−11 3.8E−10 Min 419 297 297 399 297 399 Max 116000 146000 116000 146000 116000 146000 n 180 136 192 124 204 112 (Patient) UO only Median 12100 22000 12100 22000 12100 22000 Average 25500 39700 25700 40200 25700 40200 Stdev 31400 37400 31200 38400 31200 38400 p (t-test) 0.0012 0.0013 0.0013 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n 239 76 245 70 245 70 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.70   0.68   0.65   0.72   0.71   0.64   0.71   0.70   0.64 SE   0.030   0.031   0.038   0.030   0.031   0.039   0.031   0.032   0.039 p Value 1.4E−11 6.5E−9 1.3E−4 1.1E−13 1.4E−11 3.9E−4 1.1E−11 8.0E−10 3.9E−4 nCohort Non-  173  180  239  189  192  245  201  204  245 persistent nCohort Persistent  144  136   76  128  124   70  116  112   70 Cutoff Quartile 2  6030  6020  6080  6030  6020  6080  6030  6020  6080 Sensitivity   87%   85%   91%   89%   88%   90%   91%   89%   90% Specificity   35%   33%   30%   34%   33%   29%   34%   33%   29% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity   67%   65%   66%   70%   69%   66%   70%   69%   66% Specificity   64%   62%   55%   63%   62%   54%   61%   60%   54% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity   40%   40%   38%   44%   43%   39%   43%   42%   39% Specificity   87%   86%   79%   87%   86%   79%   85%   84%   79% OR Quartile 2   3.49   2.83   4.25   4.27   3.63   3.75   4.88   4.08   3.75 p Value 2.0E−5 3.3E−4 5.9E−4 6.6E−6 4.2E−5   0.0018 6.0E−6 3.6E−5   0.0018 Lower limit of 95% CI   1.97   1.60   1.86   2.27   1.96   1.64   2.46   2.09   1.64 Upper limit of 95% CI   6.21   4.99   9.70   8.02   6.74   8.57   9.70   7.93   8.57 OR Quartile 3   3.49   3.05   2.33   3.88   3.55   2.28   3.65   3.34   2.28 p Value 1.3E−7 2.5E−6   0.0020 2.8E−8 2.0E−7   0.0036 1.9E−7 1.3E−6   0.0036 Lower limit of 95% CI   2.19   1.92   1.36   2.40   2.20   1.31   2.24   2.05   1.31 Upper limit of 95% CI   5.56   4.84   3.99   6.26   5.73   3.96   5.94   5.44   3.96 OR Quartile 4   4.63   4.08   2.33   5.35   4.77   2.33   4.32   3.89   2.33 p Value 7.2E−8 4.1E−7   0.0029 2.7E−9 2.0E−8   0.0036 8.1E−8 5.7E−7   0.0036 Lower limit of 95% CI   2.65   2.37   1.33   3.08   2.76   1.32   2.53   2.28   1.32 Upper limit of 95% CI   8.08   7.04   4.08   9.29   8.22   4.12   7.37   6.62   4.12

TABLE 8.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9450 22000 9510 28600 9680 24700 Average 17900 41300 17700 44200 19600 43900 Stdev 22400 39000 22200 39600 24300 40100 p (t-test) 1.5E−10 4.6E−13 8.4E−11 Min 419 297 297 399 297 399 Max 116000 146000 116000 146000 116000 146000 n 169 148 184 133 197 120 (Patient) sCr only Median 10800 22000 10200 27300 10800 22700 Average 18700 41200 18500 43600 20000 43700 Stdev 22700 39800 22600 40300 24100 40900 p (t-test) 1.0E−9 1.0E−11 3.2E−10 Min 419 297 297 399 297 399 Max 116000 146000 116000 146000 116000 146000 n 176 140 188 128 200 116 (Patient) UO only Median 11900 23400 11900 22700 11900 22700 Average 24400 42200 24500 42400 24500 42400 Stdev 29800 39400 29800 39900 29800 39900 p (t-test) 2.9E−5 3.3E−5 3.3E−5 Min 297 399 297 399 297 399 Max 127000 146000 127000 146000 127000 146000 n 234 81 237 78 237 78 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.70   0.68   0.67   0.73   0.71   0.66   0.71   0.70   0.66 SE   0.030   0.031   0.037   0.029   0.030   0.037   0.031   0.032   0.037 p Value 1.4E−11 4.0E−9 5.2E−6 8.0E−15 8.8E−12 1.3E−5 5.9E−12 5.9E−10 1.3E−5 nCohort Non-  169  176  234  184  188  237  197  200  237 persistent nCohort Persistent  148  140   81  133  128   78  120  116   78 Cutoff Quartile 2  6030  6020  6080  6030  6020  6080  6030  6020  6080 Sensitivity   86%   86%   91%   89%   88%   91%   90%   90%   91% Specificity   35%   34%   31%   35%   34%   30%   34%   34%   30% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity   66%   65%   68%   70%   68%   68%   70%   68%   68% Specificity   64%   62%   56%   64%   62%   56%   62%   60%   56% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity   40%   39%   40%   44%   42%   40%   42%   41%   40% Specificity   88%   86%   80%   88%   87%   80%   85%   84%   80% OR Quartile 2   3.43   3.03   4.70   4.20   3.89   4.43   4.64   4.37   4.43 p Value 2.1E−5 1.3E−4 2.3E−4 5.2E−6 1.6E−5 4.1E−4 6.2E−6 1.4E−5 4.1E−4 Lower limit of 95% CI   1.95   1.72   2.06   2.26   2.10   1.94   2.38   2.24   1.94 Upper limit of 95% CI   6.06   5.34   10.7   7.78   7.21   10.1   9.02   8.50   10.1 OR Quartile 3   3.47   3.02   2.69   4.16   3.50   2.67   3.80   3.27   2.67 p Value 1.4E−7 2.7E−6 2.7E−4 5.0E−9 2.3E−7 3.8E−4 7.0E−8 1.5E−6 3.8E−4 Lower limit of 95% CI   2.18   1.90   1.58   2.58   2.18   1.55   2.34   2.02   1.55 Upper limit of 95% CI   5.51   4.79   4.59   6.70   5.62   4.57   6.16   5.30   4.57 OR Quartile 4   4.67   4.10   2.60   5.69   4.76   2.60   4.28   3.85   2.60 p Value 8.1E−8 4.6E−7 6.4E−4 1.3E−9 2.4E−8 7.2E−4 1.0E−7 6.9E−7 7.2E−4 Lower limit of 95% CI   2.66   2.37   1.50   3.25   2.75   1.49   2.51   2.26   1.49 Upper limit of 95% CI   8.21   7.09   4.50   9.99   8.23   4.52   7.31   6.55   4.52

TABLE 8.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 350000 595000 365000 595000 377000 605000 Average 435000 599000 444000 605000 451000 613000 Stdev 256000 279000 261000 277000 260000 285000 p (t-test) 1.2E−7 5.6E−7 1.7E−6 Min 13600 105000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 206 121 222 105 238 89 (Patient) sCr only Median 350000 596000 359000 594000 375000 590000 Average 432000 602000 443000 604000 450000 607000 Stdev 255000 276000 261000 273000 261000 278000 p (t-test) 3.8E−8 5.1E−7 2.9E−6 Min 13600 105000 13600 151000 13600 154000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 208 118 223 103 236 90 (Patient) UO only Median 404000 700000 404000 765000 406000 765000 Average 469000 677000 468000 691000 470000 693000 Stdev 264000 283000 264000 283000 264000 285000 p (t-test) 2.3E−6 7.2E−7 1.1E−6 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n 281 44 283 42 285 40 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC    0.67    0.68    0.71    0.67    0.67    0.72    0.66    0.66    0.72 SE    0.032    0.032    0.046    0.033    0.033    0.047    0.035    0.035    0.048 p Value 9.0E−8 2.5E−8 7.4E−6 4.5E−7 3.3E−7 3.0E−6 3.2E−6 3.1E−6 5.1E−6 nCohort Non-   206   208   281   222   223   283   238   236   285 persistent nCohort   121   118   44   105   103   42   89   90   40 Persistent Cutoff 260000 260000 262000 260000 260000 262000 260000 260000 262000 Quartile 2 Sensitivity   85%   86%   91%   86%   86%   90%   85%   87%   90% Specificity   31%   31%   27%   30%   30%   27%   29%   30%   27% Cutoff 437000 433000 437000 437000 433000 437000 437000 433000 437000 Quartile 3 Sensitivity   66%   68%   75%   67%   67%   79%   67%   68%   80% Specificity   59%   60%   54%   58%   58%   54%   56%   57%   54% Cutoff 707000 704000 708000 707000 704000 708000 707000 704000 708000 Quartile 4 Sensitivity   39%   39%   50%   39%   39%   52%   42%   39%   52% Specificity   83%   83%   79%   82%   81%   79%   81%   80%   79% OR Quartile 2    2.58    2.70    3.77    2.59    2.79    3.55    2.39    2.74    3.33 p Value    0.0014 1.0E−3    0.014    0.0025    0.0015    0.019    0.0089    0.0031    0.027 Lower limit    1.44    1.49    1.31    1.40    1.48    1.23    1.24    1.40    1.15 of 95% CI Upper limit    4.61    4.88   10.9    4.81    5.24   10.3    4.58    5.35    9.67 of 95% CI OR Quartile 3    2.83    3.17    3.48    2.72    2.79    4.32    2.67    2.76    4.70 p Value 1.3E−5 2.0E−6 7.0E−4 5.2E−5 4.1E−5 2.1E−4 1.7E−4 9.9E−5 1.8E−4 Lower limit    1.77    1.97    1.69    1.68    1.71    1.99    1.60    1.66    2.09 of 95% CI Upper limit    4.52    5.10    7.17    4.42    4.54    9.35    4.45    4.61   10.6 of 95% CI OR Quartile 4    3.10    3.05    3.68    2.83    2.74    4.09    3.05    2.56    4.06 p Value 1.7E−5 2.2E−5 9.9E−5 8.4E−5 1.5E−4 3.7E−5 3.9E−5 5.2E−4 5.7E−5 Lower limit    1.85    1.82    1.91    1.68    1.63    2.09    1.79    1.51    2.05 of 95% CI Upper limit    5.20    5.11    7.10    4.75    4.60    7.98    5.19    4.35    8.03 of 95% CI

TABLE 8.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 338000 594000 359000 586000 371000 597000 Average 420000 596000 433000 599000 439000 609000 Stdev 248000 280000 253000 282000 253000 287000 p (t-test) 5.0E−9 7.2E−8 1.0E−7 Min 13600 105000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 187 140 204 123 219 108 (Patient) sCr only Median 341000 590000 359000 576000 371000 578000 Average 424000 593000 437000 589000 443000 594000 Stdev 253000 275000 259000 276000 259000 278000 p (t-test) 2.7E−8 1.0E−6 2.0E−6 Min 13600 105000 13600 151000 13600 154000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 192 134 205 121 217 109 (Patient) UO only Median 398000 600000 400000 607000 402000 607000 Average 462000 638000 462000 646000 463000 646000 Stdev 262000 290000 260000 293000 260000 295000 p (t-test) 3.2E−6 1.5E−6 2.5E−6 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n 260 65 263 62 265 60 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC    0.68    0.68    0.68    0.67    0.66    0.68    0.67    0.66    0.68 SE    0.030    0.031    0.040    0.031    0.032    0.040    0.033    0.033    0.041 p Value 1.3E−9 7.6E−9 6.7E−6 4.7E−8 4.9E−7 4.9E−6 1.3E−7 1.3E−6 8.7E−6 nCohort Non-   187   192   260   204   205   263   219   217   265 persistent nCohort   140   134   65   123   121   62   108   109   60 Persistent Cutoff 260000 260000 262000 260000 260000 262000 260000 260000 262000 Quartile 2 Sensitivity   86%   86%   89%   86%   86%   89%   86%   86%   88% Specificity   33%   33%   28%   32%   32%   28%   31%   31%   28% Cutoff 437000 433000 437000 437000 433000 437000 437000 433000 437000 Quartile 3 Sensitivity   66%   66%   68%   65%   64%   69%   66%   65%   70% Specificity   61%   61%   54%   59%   59%   54%   58%   58%   54% Cutoff 707000 704000 708000 707000 704000 708000 707000 704000 708000 Quartile 4 Sensitivity   38%   37%   43%   38%   36%   45%   40%   37%   45% Specificity   84%   83%   79%   83%   81%   79%   82%   81%   79% OR Quartile 2    2.98    2.96    3.30    2.92    2.84    3.08    2.73    2.80    2.93 p Value 1.5E−4 2.0E−4    0.0048 3.8E−4 5.4E−4    0.0081    0.0014    0.0011    0.011 Lower limit    1.70    1.67    1.44    1.62    1.57    1.34    1.48    1.51    1.28 of 95% CI Upper limit    5.22    5.23    7.55    5.26    5.13    7.06    5.06    5.18    6.75 of 95% CI OR Quartile 3    3.06    3.15    2.48    2.66    2.56    2.70    2.60    2.54    2.78 p Value 1.6E−6 1.1E−6    0.0019 3.6E−5 7.3E−5    0.0010 9.3E−5 1.3E−4 9.0E−4 Lower limit    1.94    1.99    1.40    1.67    1.61    1.49    1.61    1.58    1.52 of 95% CI Upper limit    4.83    5.00    4.41    4.23    4.08    4.87    4.20    4.09    5.07 of 95% CI OR Quartile 4    3.32    2.98    2.89    2.99    2.51    3.19    3.05    2.42    3.12 p Value 6.9E−6 3.5E−5 3.0E−4 3.1E−5 4.1E−4 9.7E−5 2.4E−5 7.9E−4 1.5E−4 Lower limit    1.97    1.78    1.62    1.78    1.51    1.78    1.82    1.44    1.73 of 95% CI Upper limit    5.60    4.99    5.13    5.00    4.19    5.71    5.13    4.04    5.63 of 95% CI

TABLE 8.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 323000 595000 339000 594000 342000 599000 Average 407000 598000 417000 603000 427000 610000 Stdev 244000 276000 246000 280000 249000 282000 p (t-test) 1.3E−10 6.4E−10 2.9E−9 Min 13600 105000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 176 151 190 137 205 122 (Patient) sCr only Median 326000 594000 339000 582000 342000 586000 Average 411000 598000 422000 598000 432000 601000 Stdev 248000 272000 252000 274000 255000 275000 p (t-test) 3.0E−10 6.3E−9 4.2E−8 Min 13600 105000 13600 151000 13600 154000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 182 144 194 132 207 119 (Patient) UO only Median 380000 607000 382000 641000 389000 609000 Average 453000 642000 455000 647000 456000 643000 Stdev 258000 286000 257000 289000 258000 290000 p (t-test) 9.1E−8 1.1E−7 2.6E−7 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n 249 76 253 72 254 71 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC    0.70    0.70    0.69    0.69    0.69    0.69    0.69    0.68    0.69 SE    0.029    0.030    0.037    0.030    0.031    0.038    0.031    0.032    0.038 p Value 5.1E−12 1.7E−11 1.5E−7 1.3E−10 1.1E−9 2.8E−7 1.3E−9 1.3E−8 6.6E−7 nCohort Non-   176   182   249   190   194   253   205   207   254 persistent nCohort Persistent   151   144   76   137   132   72   122   119   71 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity   87%   87%   89%   87%   87%   89%   87%   87%   89% Specificity   35%   35%   29%   34%   34%   29%   32%   32%   29% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity   66%   67%   68%   66%   66%   69%   66%   66%   69% Specificity   64%   64%   55%   61%   61%   55%   60%   59%   55% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity   38%   38%   43%   39%   37%   44%   39%   37%   44% Specificity   86%   85%   80%   85%   83%   80%   83%   82%   80% OR Quartile 2    3.56    3.48    3.53    3.36    3.41    3.24    3.15    3.32    3.18 p Value 9.8E−6 1.8E−5    0.0016 4.2E−5 4.6E−5    0.0032 1.9E−4 1.3E−4    0.0039 Lower limit of 95%    2.03    1.97    1.61    1.88    1.89    1.48    1.72    1.79    1.45 CI Upper limit of 95%    6.26    6.17    7.70    6.00    6.15    7.10    5.74    6.14    6.96 CI OR Quartile 3    3.43    3.63    2.69    3.00    3.00    2.82    2.90    2.89    2.74 p Value 1.2E−7 4.3E−8 3.6E−4 2.5E−6 3.0E−6 2.9E−4 8.0E−6 9.8E−6 4.3E−4 Lower limit of 95%    2.17    2.29    1.56    1.90    1.89    1.61    1.82    1.81    1.56 CI Upper limit of 95%    5.41    5.75    4.64    4.74    4.76    4.93    4.64    4.63    4.79 CI OR Quartile 4    3.66    3.30    3.13    3.50    2.88    3.25    3.26    2.61    3.08 p Value 2.1E−6 8.4E−6 4.8E−5 2.7E−6 5.6E−5 3.6E−5 7.4E−6 2.4E−4 8.2E−5 Lower limit of 95%    2.14    1.95    1.81    2.07    1.72    1.86    1.95    1.56    1.76 CI Upper limit of 95%    6.26    5.58    5.43    5.92    4.82    5.68    5.47    4.35    5.40 CI

TABLE 8.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 321000 590000 339000 586000 342000 595000 Average 406000 595000 417000 601000 425000 606000 Stdev 246000 274000 247000 279000 250000 280000 p (t-test) 1.9E−10 9.9E−10 3.2E−9 Min 13600 105000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 173 154 188 139 200 127 (Patient) sCr only Median 326000 594000 334000 582000 341000 590000 Average 408000 598000 417000 600000 425000 605000 Stdev 247000 272000 250000 274000 252000 275000 p (t-test) 1.5E−10 1.2E−9 3.9E−9 Min 13600 105000 13600 151000 13600 154000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 179 147 190 136 202 124 (Patient) UO only Median 377000 596000 382000 602000 389000 600000 Average 449000 632000 451000 637000 453000 634000 Stdev 257000 285000 256000 289000 257000 290000 p (t-test) 8.9E−8 1.0E−7 2.4E−7 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n 240 85 245 80 246 79 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC    0.70    0.70    0.69    0.69    0.69    0.69    0.69    0.69    0.68 SE    0.029    0.029    0.035    0.030    0.030    0.036    0.031    0.031    0.036 p Value 5.3E−12 5.7E−12 9.1E−8 1.7E−10 1.3E−10 1.8E−7 1.0E−9 7.6E−10 4.3E−7 nCohort Non-   173   179   240   188   190   245   200   202   246 persistent nCohort Persistent   154   147   85   139   136   80   127   124   79 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity   87%   87%   91%   87%   88%   90%   87%   88%   90% Specificity   36%   35%   30%   34%   34%   30%   33%   33%   30% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity   66%   67%   67%   65%   66%   68%   66%   67%   67% Specificity   64%   64%   56%   61%   62%   56%   60%   60%   55% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity   37%   37%   41%   38%   38%   42%   39%   38%   42% Specificity   86%   85%   80%   85%   84%   80%   84%   83%   80% OR Quartile 2    3.74    3.66    4.21    3.47    3.64    3.82    3.42    3.61    3.74 p Value 4.4E−6 8.6E−6 3.0E−4 2.6E−5 1.8E−5 7.6E−4 6.2E−5 4.2E−5 9.1E−4 Lower limit of 95%    2.13    2.07    1.93    1.94    2.02    1.75    1.87    1.95    1.72 CI Upper limit of 95%    6.57    6.48    9.16    6.20    6.57    8.33    6.23    6.66    8.17 CI OR Quartile 3    3.51    3.71    2.57    2.99    3.14    2.59    2.93    3.09    2.52 p Value 6.8E−8 2.5E−8 3.6E−4 2.6E−6 1.1E−6 4.4E−4 5.5E−6 2.4E−6 6.7E−4 Lower limit of 95%    2.23    2.34    1.53    1.89    1.98    1.52    1.84    1.93    1.48 CI Upper limit of 95%    5.54    5.87    4.32    4.71    4.97    4.41    4.66    4.93    4.29 CI OR Quartile 4    3.48    3.37    2.87    3.38    3.08    3.03    3.18    2.91    2.88 p Value 5.0E−6 6.7E−6 1.2E−4 5.1E−6 2.1E−5 6.4E−5 1.2E−5 4.6E−5 1.4E−4 Lower limit of 95%    2.04    1.98    1.68    2.00    1.83    1.76    1.90    1.74    1.67 CI Upper limit of 95%    5.94    5.71    4.92    5.70    5.17    5.23    5.33    4.87    4.98 CI

TABLE 8.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 314000 577000 327000 572000 339000 578000 Average 407000 590000 417000 595000 424000 602000 Stdev 248000 273000 250000 276000 252000 277000 p (t-test) 6.5E−10 2.6E−9 5.7E−9 Min 13600 105000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 169 158 183 144 196 131 (Patient) sCr only Median 321000 586000 328000 577000 340000 582000 Average 406000 595000 416000 597000 424000 602000 Stdev 246000 272000 249000 274000 252000 275000 p (t-test) 1.8E−10 1.4E−9 5.1E−9 Min 13600 105000 13600 151000 13600 154000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 175 151 186 140 198 128 (Patient) UO only Median 371000 602000 375000 607000 377000 605000 Average 442000 640000 443000 644000 444000 641000 Stdev 252000 287000 251000 289000 252000 289000 p (t-test) 2.8E−9 2.2E−9 5.8E−9 Min 13600 135000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n 235 90 237 88 238 87 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC    0.70    0.70    0.70    0.69    0.69    0.70    0.69    0.69    0.70 SE    0.029    0.029    0.034    0.030    0.030    0.034    0.031    0.031    0.035 p Value 1.4E−11 4.7E−12 2.7E−9 2.1E−10 1.2E−10 3.0E−9 9.8E−10 7.8E−10 8.1E−9 nCohort Non-   169   175   235   183   186   237   196   198   238 persistent nCohort Persistent   158   151   90   144   140   88   131   128   87 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity   87%   87%   91%   88%   87%   91%   88%   88%   91% Specificity   36%   35%   31%   35%   34%   31%   34%   33%   31% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity   66%   67%   68%   65%   66%   68%   66%   66%   68% Specificity   64%   65%   57%   62%   62%   57%   61%   61%   56% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity   36%   37%   42%   37%   37%   43%   37%   38%   43% Specificity   85%   85%   81%   84%   84%   81%   83%   83%   81% OR Quartile 2    3.68    3.59    4.62    3.76    3.56    4.45    3.65    3.50    4.37 p Value 4.3E−6 8.6E−6 1.1E−4 7.4E−6 1.8E−5 1.7E−4 2.4E−5 4.5E−5 2.0E−4 Lower limit of 95%    2.11    2.05    2.12    2.11    1.99    2.05    2.00    1.92    2.01 CI Upper limit of 95%    6.43    6.31   10.0    6.72    6.35    9.68    6.66    6.39    9.51 CI OR Quartile 3    3.50    3.68    2.74    3.03    3.10    2.79    3.06    3.04    2.71 p Value 7.0E−8 2.6E−8 1.1E−4 1.7E−6 1.2E−6 1.0E−4 2.2E−6 2.7E−6 1.6E−4 Lower limit of 95%    2.22    2.33    1.64    1.93    1.96    1.66    1.92    1.91    1.62 CI Upper limit of 95%    5.52    5.83    4.58    4.78    4.91    4.67    4.85    4.84    4.56 CI OR Quartile 4    3.25    3.38    3.17    3.09    3.07    3.33    2.95    2.89    3.17 p Value 1.5E−5 7.2E−6 2.1E−5 2.2E−5 2.3E−5 9.9E−6 3.8E−5 5.1E−5 2.3E−5 Lower limit of 95%    1.90    1.99    1.86    1.84    1.83    1.95    1.76    1.73    1.86 CI Upper limit of 95%    5.55    5.75    5.40    5.21    5.17    5.69    4.94    4.84    5.42 CI

Example 9. Use of Metalloproteinase Inhibitor 1 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 1 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 9.1 Compaarison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine outpute (UO) only, or serum creatinineor urine output RIFLE criteria Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 7870 21100 9400 20500 9450 22400 Average 15500 36800 18800 38800 19400 41100 Stdev 22400 36200 24100 38000 25100 38400 p (t-test) 1.6E−8 4.3E−8 3.9E−9 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 119 198 158 159 179 138 (Patient) sCr only Median 8850 20700 9980 20700 9560 23100 Average 16600 37100 18200 39700 19100 42000 Stdev 22300 37000 22300 39000 24000 39400 p (t-test) 3.2E−8 3.4E−9 5.1E−10 Min 475 297 419 297 399 297 Max 116000 146000 116000 146000 116000 146000 n 130 186 162 154 184 132 (Patient) UO only Median 11800 27400 12100 29500 12100 31500 Average 24600 43300 25100 45500 25300 47500 Stdev 30500 38400 30600 39800 30600 40500 p (t-test) 2.0E−5 1.9E−5 9.4E−6 Min 297 873 297 873 297 873 Max 146000 140000 146000 140000 146000 140000 n 241 74 256 59 263 52 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.73   0.70   0.68   0.68   0.68   0.68   0.70   0.70   0.69 SE   0.028   0.029   0.038   0.030   0.030   0.041   0.030   0.030  0.043 p Value 6.7E−16 7.2E−12 1.0E−6 1.7E−9 2.5E−9 7.7E−6 2.7E−11 4.0E−11 7.8E−6 nCohort Non-  119  130  241  158  162  256  179  184  263 persistent nCohort Persistent  198  186   74  159  154   59  138  132   52 Cutoff Quartile 2  6030  6020  6080  6030  6020  6080  6030  6020  6080 Sensitivity   86%   84%   92%   86%   85%   92%   88%   86%   90% Specificity   43%   38%   30%   36%   35%   29%   35%   33%   28% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity   65%   65%   70%   64%   64%   71%   69%   70%   73% Specificity   74%   71%   56%   64%   64%   55%   64%   64%   54% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity   34%   35%   39%   37%   38%   41%   40%   41%   42% Specificity   90%   89%   79%   87%   88%   79%   86%   86%   78% OR Quartile 2   4.55   3.38   4.92   3.51   3.01   4.39   3.77   3.14   3.68 p Value 3.8E−8 6.8E−6 3.8E−4 9.1E−6 8.3E−5   0.0024 1.2E−5 1.2E−4   0.0078 Lower limit of 95% CI   2.65   1.99   2.05   2.02   1.74   1.69   2.08   1.75   1.41 Upper limit of 95% CI   7.81   5.75   11.9   6.12   5.21   11.4   6.83   5.63   9.62 OR Quartile 3   5.19   4.40   3.01   3.17   3.14   2.98   3.97   4.11   3.23 p Value 1.3E−10 1.8E−9 1.1E−4 8.2E−7 1.0E−6 5.0E−4 1.1E−8 6.8E−9 4.8E−4 Lower limit of 95% CI   3.14   2.72   1.72   2.00   1.98   1.61   2.48   2.55   1.67 Upper limit of 95% CI   8.58   7.13   5.27   5.02   4.97   5.51   6.37   6.63   6.25 OR Quartile 4   4.66   4.45   2.46   3.85   4.41   2.51   4.08   4.40   2.65 p Value 5.6E−6 3.5E−6   0.0016 2.5E−6 3.3E−7   0.0026 3.8E−7 1.0E−7   0.0022 Lower limit of 95% CI   2.40   2.37   1.40   2.20   2.49   1.38   2.37   2.55   1.42 Upper limit of 95% CI   9.07   8.37   4.31   6.74   7.80   4.56   7.02   7.60   4.94

TABLE 9.2 Compaarison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine outpute (UO) only, or serum creatinineor urine output RIFLE criteria Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 7070 20500 9020 20700 9010 22400 Average 15500 36100 17600 38500 18200 40400 Stdev 22900 35900 22700 37800 24100 37900 p (t-test) 7.2E−8 1.2E−8 1.2E−9 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 112 205 147 170 165 152 (Patient) sCr only Median 8650 20500 9980 20500 9450 22000 Average 16600 36700 18300 38600 18900 39900 Stdev 22600 36700 22800 38400 24300 38400 p (t-test) 9.4E−8 3.1E−8 1.0E−8 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 126 190 154 162 169 147 (Patient) UO only Median 11600 21900 12100 21900 12100 24700 Average 24400 39900 25100 41000 25200 41900 Stdev 30500 37400 30600 38700 30600 39300 p (t-test) 1.6E−4 2.6E−4 2.0E−4 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n 222 93 238 77 245 70 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.73   0.70   0.66   0.69   0.68   0.65   0.71   0.69   0.65 SE   0.028   0.029   0.035   0.029   0.030   0.038   0.029   0.030  0.039 p V alue 4.4E−16 1.7E−11 3.2E−6 4.3E−11 3.8E−9 5.3E−5 1.1E−13 1.1E−10 7.5E−5 nCohort Non-  112  126  222  147  154  238  165  169  245 persistent nCohort Persistent  205  190   93  170  162   77  152  147   70 Cutoff Quartile 2  6030  6020  6080  6030  6020  6080  6030  6020  6080 Sensitivity   86%   84%   92%   87%   86%   92%   89%   87%   91% Specificity   45%   39%   32%   39%   36%   31%   38%   36%   30% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity   63%   64%   67%   64%   63%   66%   68%   67%   67% Specificity   74%   71%   57%   65%   64%   55%   66%   64%   55% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity   34%   34%   37%   36%   36%   36%   39%   38%   37% Specificity   90%   89%   80%   88%   87%   79%   87%   86%   78% OR Quartile 2   4.89   3.39   5.90   4.26   3.45   5.24   4.78   3.71   4.53 p Value 9.0E−9 6.1E−6 2.2E−5 3.5E−7 1.0E−5 2.2E−4 2.5E−7 8.1E−6 7.8E−4 Lower limit of 95% CI   2.85   2.00   2.60   2.44   1.99   2.18   2.64   2.09   1.88 Upper limit of 95% CI   8.41   5.76   13.4   7.44   5.99   12.6   8.66   6.60   10.9 OR Quartile 3   4.96   4.22   2.63   3.28   2.98   2.40   4.09   3.63   2.47 p Value 7.2E−10 5.6E−9 1.9E−4 4.5E−7 3.0E−6   0.0014 3.8E−9 5.6E−8   0.0015 Lower limit of 95% CI   2.98   2.60   1.58   2.07   1.88   1.40   2.56   2.28   1.41 Upper limit of 95% CI   8.26   6.85   4.36   5.20   4.70   4.11   6.54   5.79   4.31 OR Quartile 4   4.66   4.16   2.27   4.11   3.84   2.10   4.35   3.91   2.14 p Value 1.1E−5 9.6E−6   0.0027 2.0E−6 3.5E−6   0.0094 3.0E−7 1.3E−6   0.0091 Lower limit of 95% CI  2.34  2.21  1.33  2.29  2.17  1.20  2.48  2.25  1.21 Upper limit of 95% CI  9.25  7.82  3.87  7.38  6.78  3.66  7.63  6.78  3.79

TABLE 9.3 Compaarison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine outpute (UO) only, or serum creatinineor urine output RIFLE criteria Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 6980 20500 9010 20500 9010 21900 Average 15500 36000 17500 38200 18400 39200 Stdev 23000 35800 22800 37600 24500 37600 p (t-test) 9.9E−8 1.7E−8 1.4E−8 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 111 206 144 173 158 159 (Patient) sCr only Median 8510 20500 9450 20500 9450 20800 Average 16800 36400 18400 38100 19200 38700 Stdev 22700 36700 23000 38200 24700 38100 p (t-test) 1.8E−7 7.7E−8 1.1E−7 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 124 192 151 165 163 153 (Patient) UO only Median 11600 20500 11900 20900 12000 21700 Average 24900 37300 25300 38000 25500 38600 Stdev 31100 36600 31100 37300 31100 37800 p (t-test) 0.0019 0.0022 0.0021 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n 212 103 225 90 232 83 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.73   0.69   0.64   0.70   0.67   0.63   0.70   0.68   0.63 SE   0.028   0.029   0.034   0.029   0.030   0.036   0.029   0.030  0.037 p Value 6.7E−16 3.3E−11 6.2E−5 1.3E−11 5.2E−9 2.8E−4 3.1E−12 1.5E−9 4.3E−4 nCohort Non-  111  124  212  144  151  225  158  163  232 persistent nCohort Persistent  206  192  103  173  165   90  159  153   83 Cutoff Quartile 2  6030  6020  6080  6030  6020  6080  6030  6020  6080 Sensitivity   86%   84%   89%   87%   86%   89%   87%   86%   88% Specificity   45%   40%   32%   40%   37%   31%   37%   36%   30% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity   63%   63%   63%   64%   62%   63%   66%   65%   64% Specificity   74%   70%   56%   66%   64%   55%   66%   64%   55% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity   33%   34%   34%   36%   36%   34%   38%   37%   35% Specificity   90%   89%   79%   88%   87%   79%   87%   86%   78% OR Quartile 2   5.00   3.53   3.95   4.50   3.64   3.54   4.14   3.47   3.09 p Value 5.9E−9 3.2E−6 9.2E−5 1.3E−7 4.2E−6 5.4E−4 9.8E−7 1.4E−5   0.0021 Lower limit of 95% CI   2.91   2.08   1.98   2.57   2.10   1.73   2.34   1.98   1.51 Upper limit of 95% CI   8.60   6.00   7.86   7.86   6.31   7.24   7.32   6.09   6.34 OR Quartile 3   4.84   4.01   2.19   3.39   2.90   2.12   3.74   3.23   2.14 p Value 1.3E−9 1.8E−8   0.0015 2.5E−7 5.0E−6   0.0034 2.5E−8 5.9E−7   0.0040 Lower limit of 95% CI   2.91   2.47   1.35   2.13   1.84   1.28   2.35   2.04   1.27 Upper limit of 95% CI   8.05   6.50   3.55   5.38   4.58   3.51   5.96   5.12   3.58 OR Quartile 4   4.58   4.02   1.97   4.28   3.65   1.94   4.18   3.51   1.95 p Value 1.4E−5 1.6E−5   0.012 1.6E−6 8.1E−6   0.016 8.0E−7 7.6E−6   0.017 Lower limit of 95% CI   2.30   2.14   1.16   2.36   2.07   1.13   2.37   2.03   1.13 Upper limit of 95% CI   9.10   7.56   3.32   7.74   6.43   3.32   7.38   6.09   3.39

TABLE 9.4 Compaarison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine outpute (UO) only, or serum creatinineor urine output RIFLE criteria Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 7070 20500 9010 20500 9010 21700 Average 15600 35900 17600 38000 18500 38800 Stdev 23000 35800 22900 37600 24600 37500 p (t-test) 1.5E−7 2.8E−8 2.9E−8 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 110 207 143 174 156 161 (Patient) sCr only Median 8510 20500 9980 19900 9560 20700 Average 16800 36400 18500 37900 19300 38500 Stdev 22700 36700 23000 38200 24700 38100 p (t-test) 1.8E−7 1.3E−7 1.8E−7 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n 124 192 150 166 162 154 (Patient) UO only Median 11800 20500 11900 20500 11900 21300 Average 25100 36400 25300 37300 25500 37600 Stdev 31300 36300 31300 36700 31400 36900 p (t-test) 0.0044 0.0034 0.0038 Min 297 399 297 399 297 399 Max 146000 140000 146000 140000 146000 140000 n 208 107 220 95 226 89 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.72   0.69   0.62   0.69   0.67   0.63   0.70   0.68   0.63 SE   0.028   0.029   0.034   0.029   0.030   0.035   0.029   0.030   0.036 p Value 2.7E−15 3.3E−11 2.3E−4 5.1E−11 1.6E−8 2.9E−4 8.7E−12 5.0E−9 4.1E−4 nCohort  110  124  208  143  150  220  156  162  226 Non- persistent nCohort  207  192  107  174  166   95  161  154   89 Persistent Cutoff 6030  6020  6080  6030  3020  6080  6030  6020  6080 Quartile 2 Sensitivity   86%   84%   88%   87%   86%   88%   87%   86%   88% Specificity   45%   40%   32%   39%   37%   31%   37%   35%   30% Cutoff 14200 14100 14200 14200 14100 14200 14200 14100 14200 Quartile 3 Sensitivity   63%   63%   62%   63%   62%   63%   66%   64%   64% Specificity   74%   70%   56%   66%   63%   55%   66%   64%   55% Cutoff 35900 35400 36300 35900 35400 36300 35900 35400 36300 Quartile 4 Sensitivity   33%   34%   34%   36%   36%   35%   37%   36%   35% Specificity   90%   89%   79%   88%   87%   79%   87%   86%   79% OR   4.74   3.53   3.36   4.23   3.43   3.42   3.95   3.26   3.05 Quartile 2 p Value 1.6E−8 3.2E−6 2.5E−4 3.2E−7 9.6E−6 4.9E−4 1.7E−6 3.0E−5   0.0016 Lower limit   2.76   2.08   1.76   2.43   1.99   1.71   2.25   1.87   1.53 of 95% CI Upper limit   8.13   6.00   6.43   7.34   5.91   6.81   6.92   5.67   6.10 of 95% CI OR   4.72   4.01   2.03   3.30   2.82   2.13   3.75   3.14   2.20 Quartile 3 p Value 2.4E−9 1.8E−8   0.0036 4.4E−7 8.4E−6   0.0027 2.5E−8 1.0E−6   0.0022 Lower limit   2.83   2.47   1.26   2.08   1.79   1.30   2.35   1.98   1.33 of 95% CI Upper limit   7.85   6.50   3.27   5.24   4.46   3.50   5.96   4.97   3.66 of 95% CI OR   4.50   4.02   1.95   4.21   3.58   2.01   4.04   3.45   1.98 Quartile 4 p Value 1.8E−5 1.6E−5   0.013 2.1E−6 1.1E−5   0.010 1.4E−6 1.0E−5   0.013 Lower limit   2.26   2.14   1.15   2.32   2.03   1.18   2.29   1.99   1.15 of 95% CI Upper limit   8.94   7.56   3.28   7.61   6.33   3.43   7.13   5.99   3.40 of 95% CI

TABLE 9.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 6950 20500 8810 20500 8870 21300 Average 14700 35900 17100 37900 18100 38500 Stdev 21600 35900 22000 37600 24000 37500 p (t-test) 4.5E−8 1.7E−8 3.0E−8 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 106 211 138 179 150 167 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 8210 20500 9380 20500 9380 21100 Average 15900 36600 17700 38100 18600 38700 Stdev 21400 36800 22000 38200 23900 38000 p (t-test) 3.8E−8 2.7E−8 4.1E−8 Min 475 297 419 297 419 297 Max 116000 146000 116000 146000 116000 146000 n (Patient) 121 195 146 170 158 158 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 11200 21300 11600 21300 11800 21500 Average 23600 38400 24100 39000 24400 39000 Stdev 29300 38100 29600 38400 29800 38700 p (t-test) 1.5E−4 1.9E−4 2.9E−4 Min 297 399 297 399 297 399 Max 127000 146000 127000 146000 127000 146000 n (Patient) 202 113 212 103 217 98 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.70 0.65 0.69 0.68 0.65 0.70 0.69 0.64 SE 0.028 0.029 0.033 0.029 0.030 0.034 0.029 0.030 0.035 p Value  2.2E−16  1.9E−12 7.3E−6  2.6E−11 1.1E−9 1.7E−5  1.0E−11  3.7E−10 3.5E−5 nCohort Non- 106 121 202 138 146 212 150 158 217 persistent nCohort Persistent 211 195 113 179 170 103 167 158 98 Cutoff Quartile 2 6030 6020 6080 6030 6020 6080 6030 6020 6080 Sensitivity 85% 85% 88% 87% 86% 89% 87% 86% 89% Specificity 45% 40% 33% 40% 38% 32% 38% 36% 31% Cutoff Quartile 3 14200 14100 14200 14200 14100 14200 14200 14100 14200 Sensitivity 63% 63% 64% 63% 62% 64% 65% 65% 64% Specificity 75% 71% 57% 66% 64% 57% 66% 65% 56% Cutoff Quartile 4 35900 35400 36300 35900 35400 36300 35900 35400 36300 Sensitivity 34% 34% 35% 36% 36% 35% 37% 37% 35% Specificity 92% 90% 80% 88% 88% 80% 87% 87% 79% OR Quartile 2 4.81 3.74 3.73 4.28 3.68 3.95 4.04 3.49 3.61 p Value 1.2E−8 1.2E−6 6.8E−5 2.1E−7 3.0E−6 9.2E−5 8.8E−7 1.0E−5 2.6E−4 Lower limit of 95% CI 2.80 2.20 1.95 2.47 2.13 1.98 2.32 2.00 1.81 Upper limit of 95% CI 8.25 6.37 7.14 7.41 6.35 7.86 7.05 6.08 7.19 OR Quartile 3 4.89 4.20 2.37 3.24 2.99 2.33 3.55 3.32 2.31 p Value 2.0E−9 8.9E−9 3.6E−4 7.1E−7 2.9E−6 6.5E−4 8.2E−8 3.4E−7 8.6E−4 Lower limit of 95% CI 2.91 2.57 1.47 2.03 1.89 1.43 2.24 2.09 1.41 Upper limit of 95% CI 8.21 6.84 3.81 5.15 4.74 3.78 5.65 5.26 3.78 OR Quartile 4 5.47 4.75 2.13 4.24 3.98 2.11 3.97 3.78 2.03 p Value 6.9E−6 4.4E−6 0.0042 2.7E−6 3.6E−6 0.0053 2.6E−6 3.4E−6 0.0088 Lower limit of 95% CI 2.61 2.44 1.27 2.32 2.22 1.25 2.23 2.16 1.20 Upper limit of 95% CI 11.5 9.25 3.59 7.76 7.14 3.57 7.05 6.64 3.45

TABLE 9.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 279000 564000 321000 571000 342000 577000 Average 363000 566000 410000 572000 418000 589000 Stdev 236000 270000 253000 274000 249000 279000 p (t-test) 5.9E−11 6.3E−8 1.3E−8 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 114 213 155 172 179 148 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 280000 567000 314000 586000 340000 594000 Average 365000 572000 394000 586000 409000 600000 Stdev 235000 268000 237000 276000 244000 276000 p (t-test) 9.2E−12 8.8E−11 1.6E−10 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 124 202 157 169 182 144 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 400000 595000 402000 660000 400000 693000 Average 457000 619000 461000 643000 460000 673000 Stdev 260000 289000 261000 288000 261000 280000 p (t-test) 3.7E−6 1.5E−6 8.5E−8 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 245 80 261 64 269 56 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.73 0.66 0.68 0.70 0.68 0.68 0.70 0.71 SE 0.028 0.028 0.037 0.029 0.029 0.040 0.030 0.029 0.041 p Value  1.1E−15  2.2E−16 8.6E−6 2.5E−9  1.1E−12 4.0E−6 2.2E−9  8.8E−12 2.1E−7 nCohort Non- 114 124 245 155 157 261 179 182 269 persistent nCohort Persistent 213 202 80 172 169 64 148 144 56 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 85% 86% 85% 85% 86% 89% 86% 88% 91% Specificity 43% 43% 28% 36% 37% 28% 35% 35% 28% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 61% 62% 65% 62% 63% 69% 63% 65% 75% Specificity 70% 70% 55% 63% 64% 54% 60% 62% 55% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 33% 34% 44% 34% 37% 47% 38% 40% 50% Specificity 90% 90% 81% 85% 87% 80% 85% 86% 80% OR Quartile 2 4.11 4.45 2.22 3.18 3.54 3.22 3.39 3.80 4.02 p Value 1.3E−7 3.4E−8 0.020 2.0E−5 4.5E−6 0.0057 2.1E−5 6.6E−6 0.0044 Lower limit of 95% 2.43 2.62 1.13 1.87 2.06 1.41 1.93 2.13 1.54 CI Upper limit of 95% 6.95 7.57 4.36 5.40 6.07 7.39 5.95 6.78 10.4 CI OR Quartile 3 3.69 3.90 2.24 2.69 3.11 2.63 2.57 2.92 3.67 p Value 1.5E−7 2.5E−8 0.0025 1.5E−5 8.6E−7 0.0012 3.6E−5 3.7E−6 9.0E−5 Lower limit of 95% 2.27 2.42 1.33 1.72 1.98 1.47 1.64 1.85 1.91 CI Upper limit of 95% 6.00 6.29 3.79 4.20 4.89 4.70 4.03 4.59 7.03 CI OR Quartile 4 4.68 4.43 3.28 3.00 3.97 3.55 3.58 4.11 3.98 p Value 9.7E−6 5.9E−6 1.9E−5 7.5E−5 1.7E−6 1.7E−5 2.6E−6 2.6E−7 7.0E−6 Lower limit of 95% 2.36 2.33 1.90 1.74 2.26 1.99 2.10 2.40 2.18 CI Upper limit of 95% 9.28 8.43 5.65 5.16 6.98 6.32 6.10 7.05 7.28 CI

TABLE 9.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 272000 565000 300000 573000 314000 578000 Average 348000 568000 389000 579000 397000 594000 Stdev 224000 270000 237000 276000 237000 278000 p (t-test) 2.1E−12 1.9E−10 3.2E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 108 219 144 183 164 163 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 277000 571000 299000 578000 314000 582000 Average 356000 574000 385000 585000 399000 592000 Stdev 228000 268000 236000 273000 241000 274000 p (t-test) 7.4E−13 1.2E−11 6.4E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 120 206 149 177 166 160 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Median 375000 590000 382000 602000 377000 609000 Average 446000 612000 451000 629000 451000 644000 Stdev 254000 290000 256000 290000 256000 287000 p (t-test) 3.7E−7 2.3E−7 4.7E−8 Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 225 100 241 84 248 77 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.74 0.74 0.67 0.70 0.72 0.68 0.71 0.71 0.69 SE 0.027 0.027 0.034 0.028 0.028 0.036 0.029 0.029 0.037 p Value 0 0 6.4E−7  5.3E−13  1.2E−14 4.7E−7  2.8E−13  5.1E−13 9.9E−8 nCohort Non- 108 120 225 144 149 241 164 166 248 persistent nCohort Persistent 219 206 100 183 177 84 163 160 77 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 85% 86% 86% 86% 86% 89% 88% 88% 90% Specificity 45% 44% 30% 39% 39% 30% 38% 37% 29% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 61% 63% 63% 62% 64% 65% 63% 64% 69% Specificity 72% 72% 56% 65% 66% 55% 63% 63% 56% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 33% 34% 42% 35% 36% 44% 37% 37% 45% Specificity 92% 90% 82% 88% 87% 81% 87% 86% 81% OR Quartile 2 4.68 4.83 2.60 3.84 4.06 3.55 4.35 4.17 3.60 p Value 1.1E−8 7.0E−9 0.0030 7.5E−7 4.0E−7 8.5E−4 3.4E−7 7.0E−7 0.0013 Lower limit of 95% 2.76 2.83 1.38 2.25 2.36 1.69 2.47 2.37 1.65 CI Upper limit of 95% 7.95 8.23 4.91 6.55 6.99 7.47 7.64 7.34 7.86 CI OR Quartile 3 4.10 4.24 2.13 3.11 3.50 2.34 2.90 3.03 2.77 p Value 3.4E−8 6.3E−9 0.0022 1.1E−6 8.4E−8 0.0013 3.4E−6 1.5E−6 2.4E−4 Lower limit of 95% 2.48 2.60 1.31 1.97 2.21 1.39 1.85 1.93 1.61 CI Upper limit of 95% 6.77 6.90 3.45 4.90 5.53 3.91 4.54 4.75 4.77 CI OR Quartile 4 5.50 4.63 3.35 3.76 3.78 3.43 4.07 3.63 3.56 p Value 6.0E−6 5.7E−6 6.1E−6 7.4E−6 5.0E−6 7.4E−6 7.8E−7 3.5E−6 5.8E−6 Lower limit of 95% 2.63 2.39 1.98 2.11 2.14 2.00 2.33 2.11 2.06 CI Upper limit of 95% 11.5 8.98 5.66 6.72 6.69 5.88 7.11 6.26 6.17 CI

Table 9.8: Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria.

TABLE 9.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort sCr or UO Median 270000 565000 298000 573000 303000 578000 Average 345000 569000 386000 579000 394000 590000 Stdev 223000 270000 238000 275000 238000 276000 p (t-test) 9.8E−13 1.2E−10 3.2E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 107 220 142 185 158 169 sCr only Median 275000 574000 295000 582000 300000 590000 Average 349000 576000 379000 587000 388000 595000 Stdev 223000 268000 233000 271000 235000 273000 p (t-tesc) 7.8E−14 2.2E−12 2.0E−12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 118 208 146 180 160 166 UO only Median 371000 595000 373000 598000 371000 602000 Average 440000 609000 446000 620000 447000 633000 Stdev 253000 286000 256000 285000 256000 283000 p (t-test) 1.1E−7  1.5E−7  3.7E−8  Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 216 109 230 95 237 88 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.75 0.75 0.67 0.71 0.72 0.68 0.71 0.72 0.69 SE 0.027 0.027 0.033 0.028 0.028 0.034 0.028 0.028 0.035 p Value 0 0 1.1E−7  1.7E−13  4.4E−16 1.7E−7  8.4E−14  2.0E−15 4.2E−8 nCohort Non- 107 118 216 142 146 230 158 160 237 persistent nCohort Persistent 220 208 109 185 180 95 169 166 88 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 85% 86% 86% 86% 87% 88% 88% 88% 89% Specificity 46% 45% 31% 39% 40% 30% 39% 39% 30% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 61% 63% 63% 62% 64% 65% 63% 64% 68% Specificity 73% 73% 56% 65% 67% 56% 63% 64% 57% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 33% 34% 40% 35% 36% 41% 36% 37% 42% Specificity 92% 91% 82% 87% 88% 81% 87% 87% 81% OR Quartile 2 4.79 5.03 2.76 3.98 4.28 3.34 4.43 4.62 3.34 p Value 7.3E−9 3.1E−9 0.0013 3.9E−7 1.5E−7 5.9E−4 1.7E−7 1.1E−7 9.5E−4 Lower limit of 95% 2.82 2.95 1.49 2.33 2.49 1.68 2.54 2.62 1.63 CI Upper limit of 95% 8.14 8.59 5.11 6.79 7.37 6.65 7.74 8.13 6.82 CI OR Quartile 3 4.27 4.57 2.24 3.12 3.61 2.40 2.90 3.19 2.79 p Value 1.8E−8 1.6E−9 8.5E−4 1.0E−6 4.5E−8 5.4E−4 3.4E−6 5.0E−7 1.0E−4 Lower limit of 95% 2.58 2.79 1.39 1.98 2.28 1.46 1.85 2.03 1.66 CI Upper limit of 95% 7.08 7.49 3.59 4.92 5.72 3.94 4.55 5.02 4.67 CI OR Quartile 4 5.41 5.04 3.17 3.64 3.92 3.03 3.68 3.85 3.10 p Value 7.4E−6 3.5E−6 1.3E−5 1.2E−5 3.9E−6 3.7E−5 4.3E−6 2.1E−6 3.3E−5 Lower limit of 95% 2.58 2.54 1.89 2.04 2.20 1.79 2.11 2.20 1.82 CI Upper limit of 95% 11.3 9.99 5.33 6.51 7.01 5.13 6.43 6.71 5.28 CI

Table 9.9: Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria.

TABLE 9.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort sCr or UO Median 272000 565000 296000 568000 301000 574000 Average 346000 567000 386000 577000 393000 587000 Stdev 224000 270000 239000 274000 240000 275000 p (t-test) 2.2E−12 1.7E−10 6.5E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 106 221 140 187 155 172 sCr only Median 275000 574000 293000 577000 300000 582000 Average 349000 576000 378000 585000 388000 593000 Stdev 223000 268000 234000 271000 236000 272000 p (t-test) 7.8E-14 3.0E-12 3.0E-12 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 118 208 144 182 158 168 UO only Median 375000 577000 375000 590000 375000 595000 Average 443000 597000 445000 612000 445000 620000 Stdev 255000 286000 255000 286000 256000 284000 p (t-test) 1.0E-6  2.6E-7  1.1E-7  Min 13600 135000 13600 135000 13600 135000 Max 1120000 1090000 1120000 1090000 1120000 1090000 n (Patient) 211 114 223 102 229 96 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.74 0.75 0.66 0.71 0.72 0.67 0.71 0.72 0.68 SE 0.027 0.027 0.033 0.028 0.028 0.033 0.028 0.028 0.034 p Value 0 0 9.4E-7  1.7E-13  4.4E-16 2.2E-7  1.2E-13  2.2E-15 8.9E-8 nCohort Non- 106 118 211 140 144 223 155 158 229 persistent nCohort Persistent 221 208 114 187 182 102 172 168 96 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 85% 86% 86% 86% 87% 89% 88% 88% 90% Specificity 45% 45% 31% 40% 40% 31% 39% 39% 31% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 61% 63% 61% 62% 64% 64% 62% 64% 66% Specificity 73% 73% 56% 66% 67% 56% 63% 65% 56% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 33% 34% 39% 34% 35% 39% 35% 36% 40% Specificity 92% 91% 82% 87% 88% 81% 86% 87% 81% OR Quartile 2 4.55 5.03 2.73 4.13 4.44 3.78 4.67 4.78 3.86 p Value 2.0E-8 3.1E-9 0.0011 2.0E-7 7.6E-8 1.4E-4 6.6E-8 6.0E-8 2.0E-4 Lower limit of 95% 2.68 2.95 1.49 2.42 2.58 1.90 2.67 2.71 1.90 CI Upper limit of 95% 7.73 8.59 4.99 7.05 7.64 7.52 8.16 8.41 7.88 CI OR Quartile 3 4.17 4.57 2.02 3.13 3.63 2.24 2.83 3.19 2.46 p Value 3.1E-8 1.6E-9 0.0031 9.9E-7 4.3E-8 0.0010 5.6E-6 5.0E-7 3.6E-4 Lower limit of 95% 2.51 2.79 1.27 1.98 2.29 1.38 1.81 2.03 1.50 CI Upper limit of 95% 6.91 7.49 3.21 4.95 5.75 3.63 4.43 5.03 4.04 CI OR Quartile 4 5.32 5.04 2.86 3.53 3.80 2.78 3.51 3.72 2.75 p Value 9.2E-6 3.5E-6 6.3E-5 2.0E-5 6.6E-6 1.2E-4 9.8E-6 3.7E-6 1.5E-4 Lower limit of 95% 2.54 2.54 1.71 1.97 2.13 1.65 2.01 2.13 1.63 CI Upper limit of 95% 11.1 9.99 4.79 6.30 6.78 4.68 6.11 6.49 4.66 CI

TABLE 9.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort Non-persistent Cohort Persistent Cohort sCr or UO Median 272000 564000 293000 565000 294000 571000 Average 347000 562000 383000 575000 389000 585000 Stdev 227000 271000 238000 274000 239000 274000 p (t-test) 1.6E−11 1.9E−10 4.2E−11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 102 225 135 192 149 178 sCr only Median 275000 568000 293000 571000 303000 574000 Average 351000 571000 381000 579000 390000 586000 Stdev 225000 269000 236000 272000 238000 273000 p (t-test) 8.4E-13 3.2E-11 3.4E-11 Min 13600 105000 13600 105000 13600 105000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 115 211 140 186 154 172 UO only Median 369000 576000 363000 595000 367000 595000 Average 436000 601000 434000 618000 436000 623000 Stdev 250000 288000 249000 286000 250000 285000 p (t-test) 1.1E-7  5.6E-9  4.2E-9  Min 13600 135000 13600 135000 13600 135000 Max 1040000 1120000 1040000 1120000 1040000 1120000 n (Patient) 204 121 214 111 219 106 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.74 0.74 0.67 0.71 0.72 0.69 0.71 0.71 0.69 SE 0.028 0.027 0.032 0.028 0.028 0.032 0.028 0.028 0.032 p Value 0 0 8.0E-8  1.3E-13  1.4E-14 3.1E-9  3.4E-14  6.5E-14 2.0E-9 nCohort 102 115 204 135 140 214 149 154 219 Non-persistent nCohort Persistent 225 211 121 192 186 111 178 172 106 Cutoff Quartile 2 260000 260000 262000 260000 260000 262000 260000 260000 262000 Sensitivity 84% 85% 87% 86% 86% 90% 88% 87% 91% Specificity 45% 44% 32% 41% 40% 33% 40% 39% 32% Cutoff Quartile 3 437000 433000 437000 437000 433000 437000 437000 433000 437000 Sensitivity 60% 62% 62% 62% 63% 65% 62% 63% 66% Specificity 73% 72% 57% 67% 67% 57% 64% 64% 58% Cutoff Quartile 4 707000 704000 708000 707000 704000 708000 707000 704000 708000 Sensitivity 32% 34% 39% 34% 34% 41% 35% 35% 41% Specificity 91% 90% 83% 87% 87% 83% 87% 86% 82% OR Quartile 2 4.31 4.63 3.07 4.20 4.10 4.42 4.78 4.35 4.61 p Value 5.9E-8 1.4E-8 2.7E-4 1.3E-7 2.3E-7 2.1E-5 3.0E-8 1.8E-7 2.5E-5 Lower limit of 95% 2.54 2.72 1.68 2.47 2.40 2.23 2.75 2.50 2.26 CI Upper limit of 95% 7.31 7.86 5.61 7.15 7.00 8.77 8.32 7.56 9.37 CI OR Quartile 3 4.04 4.25 2.15 3.26 3.47 2.50 3.00 3.04 2.63 p Value 8.3E-8 9.3E-9 0.0011 5.2E-7 1.3E-7 1.6E-4 1.9E-6 1.4E-6 8.5E-5 Lower limit of 95% 2.42 2.59 1.36 2.06 2.18 1.55 1.91 1.93 1.63 CI Upper limit of 95% 6.73 6.96 3.40 5.17 5.50 4.01 4.72 4.77 4.27 CI OR Quartile 4 4.96 4.79 3.07 3.55 3.56 3.26 3.75 3.48 3.15 p Value 2.1E-5 7.0E-6 2.1E-5 2.5E-5 1.8E-5 7.9E-6 5.8E-6 1.1E-5 1.5E-5 Lower limit of 95% 2.37 2.42 1.83 1.97 1.99 1.94 2.12 2.00 1.87 CI Upper limit of 95% 10.4 9.50 5.14 6.41 6.35 5.48 6.64 6.07 5.30 CI

Example 10. Use of Metalloproteinase Inhibitor 2 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 2 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 10.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort sCr or UO Median 9920 16000 10100 15400 10000 15100 Average 12700 33200 13500 32200 13000 31900 Stdev 10100 58100 10900 57100 10500 56600 p (t-test) 0.0084 0.027 0.034 Min 200 658 200 658 200 658 Max 42400 465000 42400 465000 38600 465000 n (Patient) 57 260 46 271 41 276 sCr only Median 12000 14800 12000 14800 11600 14800 Average 16600 33200 16600 33100 15800 33100 Stdev 13400 60100 13600 59900 12500 59400 p (t-test) 0.017 0.020 0.016 Min 200 658 200 658 200 658 Max 63200 465000 63200 465000 63200 465000 n (Patient) 76 240 74 242 70 246 UO only Median 10600 21700 10300 21200 10100 19600 Average 19200 46300 19200 42300 19300 39700 Stdev 38500 68200 40400 63900 42200 61000 p (t-test) 9.9E-6 1.1E-4 6.3E-4 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 194 121 173 142 156 159 Recovery Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.67 0.58 0.74 0.65 0.57 0.73 0.66 0.59 0.72 SE 0.036 0.037 0.030 0.040 0.037 0.029 0.042 0.037 0.029 p Value 2.8E-6 0.037  6.7E-16 2.7E-4 0.042  5.1E-15 1.6E-4 0.018  1.6E-14 nCohort Recovered 57 76 194 46 74 173 41 70 156 nCohort Non-recovered 260 240 121 271 242 142 276 246 159 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 79% 77% 93% 78% 77% 91% 78% 77% 89% Specificity 42% 30% 37% 41% 31% 38% 44% 33% 39% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 54% 52% 69% 53% 52% 67% 53% 52% 66% Specificity 68% 57% 62% 67% 57% 64% 68% 57% 66% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 28% 28% 40% 27% 28% 37% 27% 28% 35% Specificity 89% 84% 85% 87% 84% 85% 88% 86% 85% OR Quartile 2 2.71 1.43 8.15 2.47 1.50 6.12 2.76 1.66 5.03 p Value 0.0012 0.23 1.1E-7 0.0066 0.17 4.2E-8 0.0034 0.087 6.8E-8 Lower limit of 95% CI 1.48 0.803 3.76 1.29 0.842 3.20 1.40 0.929 2.80 Upper limit of 95% CI 4.96 2.53 17.7 4.76 2.66 11.7 5.44 2.97 9.04 OR Quartile 3 2.57 1.42 3.68 2.34 1.43 3.53 2.42 1.45 3.78 p Value 0.0024 0.19 1.2E-7 0.012 0.19 1.2E-7 0.013 0.18 2.3E-8 Lower limit of 95% CI 1.40 0.842 2.27 1.21 0.844 2.21 1.20 0.847 2.37 Upper limit of 95% CI 4.72 2.38 5.97 4.54 2.41 5.63 4.87 2.47 6.02 OR Quartile 4 3.38 2.07 3.72 2.50 1.98 3.37 2.69 2.34 2.91 p Value 0.0071 0.036 1.3E-6 0.045 0.049 9.8E-6 0.046 0.022 1.2E-4 Lower limit of 95% CI 1.39 1.05 2.19 1.02 1.00 1.97 1.02 1.13 1.69 Upper limit of 95% CI 8.22 4.07 6.33 6.15 3.90 5.77 7.10 4.83 5.01

TABLE 10.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 10600 18100 10900 17200 10600 16100 Average 13600 35800 14200 34600 13700 34100 Stdev 11500 61500 12100 60300 12000 59500 p (t-test) 8.0E−4 0.0031 0.0045 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 90 227 79 238 71 246 sCr only Median 12000 16000 12000 16000 11400 16600 Average 16700 35900 16700 35700 16000 35600 Stdev 14200 63700 14300 63400 13600 62800 p (t-test) 0.0023 0.0028 0.0022 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 106 211 104 213 99 218 UO only Median 11400 21700 11200 21100 10900 19500 Average 20000 49400 19900 45100 19900 42300 Stdev 37100 73200 38400 68700 39800 65300 p (t-test) 3.4E−6 3.9E−5 2.0E−4 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 212 103 194 121 178 137 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.59 0.71 0.64 0.59 0.69 0.65 0.60 0.69 SE 0.032 0.033 0.032 0.034 0.033 0.031 0.035 0.033 0.030 p Value 5.3E−7 0.0065 7.9E−11 3.3E−5 0.0075 6.2E−10 9.4E−6 0.0018 7.6E−10 nCohort Recovered 90 106 212 79 104 194 71 99 178 nCohort Non- 227 211 103 238 213 121 246 218 137 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 79% 77% 90% 78% 77% 88% 78% 78% 87% Specificity 34% 28% 33% 34% 29% 34% 37% 30% 34% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 56% 54% 68% 55% 54% 65% 54% 54% 65% Specificity 66% 57% 58% 65% 57% 59% 65% 58% 61% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 31% 30% 41% 30% 30% 38% 30% 31% 36% Specificity 90% 85% 83% 89% 85% 83% 90% 87% 83% OR Quartile 2 1.96 1.31 4.49 1.86 1.36 3.85 2.10 1.50 3.45 p Value 0.014 0.32 3.7E−5 0.029 0.26 2.9E−5 0.011 0.14 3.3E−5 Lower limit of 95% 1.14 0.768 2.20 1.06 0.798 2.05 1.19 0.879 1.92 CI Upper limit of 95% CI 3.36 2.22 9.15 3.24 2.31 7.24 3.72 2.56 6.18 OR Quartile 3 2.46 1.50 2.99 2.23 1.51 2.74 2.20 1.57 2.93 p Value 5.1E−4 0.089 1.5E−5 0.0029 0.087 2.8E−5 0.0048 0.064 5.3E−6 Lower limit of 95% 1.48 0.940 1.82 1.32 0.942 1.71 1.27 0.973 1.84 CI Upper limit of 95% CI 4.09 2.41 4.91 3.78 2.42 4.39 3.81 2.54 4.65 OR Quartile 4 4.10 2.45 3.26 3.31 2.36 2.99 3.86 2.94 2.75 p Value 2.0E−4 0.0039 1.2E−5 0.0017 0.0056 4.2E−5 0.0014 0.0012 1.6E−4 Lower limit of 95% 1.95 1.33 1.92 1.57 1.29 1.77 1.69 1.53 1.62 CI Upper limit of 95% CI 8.62 4.50 5.53 6.98 4.34 5.06 8.82 5.62 4.65

TABLE 10.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11300 18600 11300 18100 11000 17900 Average 15000 37600 15200 36800 14600 36000 Stdev 13100 64600 13400 63700 12800 62400 p (t-test) 2.5E−4 5.9E−4 9.1E−4 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 114 203 107 210 97 220 sCr only Median 11900 18000 12000 17900 11600 17900 Average 16800 38000 17000 37500 16400 37400 Stdev 14800 66600 14900 66100 14300 65400 p (t-test) 4.9E−4 7.4E−4 6.3E−4 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 127 190 124 193 119 198 UO only Median 11600 21500 11500 21400 11300 20600 Average 20700 48500 20800 45900 20100 44000 Stdev 37700 73600 38500 70900 38900 67800 p (t-test) 1.3E−5 5.7E−5 8.9E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 214 101 204 111 190 125 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.61 0.69 0.64 0.60 0.68 0.64 0.61 0.68 SE 0.031 0.032 0.033 0.032 0.032 0.033 0.032 0.032 0.031 p Value 2.5E−6 8.2E−4 1.7E−8 1.3E−5 0.0016 4.5E−8 6.7E−6 5.3E−4 7.1E−9 nCohort Recovered 114 127 214 107 124 204 97 119 190 nCohort Non- 203 190 101 210 193 111 220 198 125 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 80% 79% 87% 80% 79% 86% 80% 79% 86% Specificity 34% 31% 31% 35% 31% 31% 36% 32% 33% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 57% 55% 65% 57% 55% 64% 56% 55% 64% Specificity 62% 57% 57% 63% 57% 57% 63% 58% 59% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 33% 32% 41% 31% 32% 39% 31% 32% 38% Specificity 88% 85% 82% 87% 85% 82% 89% 87% 83% OR Quartile 2 2.12 1.78 3.02 2.11 1.76 2.93 2.26 1.80 3.08 p Value 0.0046 0.028 8.7E−4 0.0050 0.032 6.8E−4 0.0026 0.026 2.1E−4 Lower limit of 95% 1.26 1.06 1.57 1.25 1.05 1.57 1.33 1.07 1.70 CI Upper limit of 95% CI 3.56 2.98 5.79 3.57 2.94 5.44 3.84 3.01 5.58 OR Quartile 3 2.20 1.67 2.50 2.19 1.63 2.39 2.15 1.69 2.55 p Value 9.9E−4 0.027 2.5E−4 0.0013 0.035 3.5E−4 0.0022 0.025 8.0E−5 Lower limit of 95% 1.38 1.06 1.53 1.36 1.04 1.48 1.32 1.07 1.60 CI Upper limit of 95% CI 3.52 2.63 4.09 3.53 2.57 3.84 3.51 2.68 4.07 OR Quartile 4 3.44 2.69 3.16 3.04 2.55 2.95 3.57 3.07 2.98 p Value 1.3E−4 7.5E−4 2.0E−5 5.7E−4 0.0014 5.3E−5 2.9E−4 2.7E−4 4.7E−5 Lower limit of 95% 1.83 1.51 1.86 1.62 1.44 1.75 1.79 1.68 1.76 CI Upper limit of 95% CI 6.47 4.78 5.38 5.73 4.54 4.99 7.12 5.63 5.03

TABLE 10.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11400 18600 11300 18400 10900 18200 Average 17300 38500 17300 37400 17000 36500 Stdev 22300 66200 22900 64900 23500 63300 p (t-test) 4.3E−4 9.4E−4 0.0017 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 134 183 125 192 114 203 sCr only Median 11800 18200 11800 18100 11600 18100 Average 18000 38600 18100 38300 17800 38200 Stdev 22400 67300 22600 66900 22400 66500 p (t-test) 6.0E−4 7.8E−4 6.9E−4 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 140 177 138 179 135 182 UO only Median 11600 21400 11400 21500 11400 20600 Average 20700 49000 20600 46900 20200 44700 Stdev 37500 74400 38100 71700 38500 68700 p (t-test) 1.0E−5 2.8E−5 6.8E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 216 99 207 108 194 121 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.62 0.68 0.64 0.62 0.68 0.65 0.62 0.67 SE 0.031 0.031 0.034 0.031 0.031 0.033 0.031 0.031 0.032 p Value 3.5E−6 9.1E−5 9.1E−8 5.6E−6 1.4E−4 4.5E−8 2.0E−6 7.2E−5 4.2E−8 nCohort Recovered 134 140 216 125 138 207 114 135 194 nCohort Non- 183 177 99 192 179 108 203 182 121 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 82% 82% 86% 82% 82% 86% 82% 81% 85% Specificity 34% 34% 30% 35% 33% 31% 37% 33% 31% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 57% 56% 65% 57% 56% 65% 57% 56% 64% Specificity 60% 57% 56% 61% 57% 57% 61% 58% 58% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 33% 33% 41% 32% 33% 40% 32% 34% 38% Specificity 86% 85% 82% 86% 85% 83% 86% 86% 83% OR Quartile 2 2.38 2.29 2.61 2.44 2.21 2.77 2.62 2.18 2.62 p Value 0.0011 0.0018 0.0031 7.6E−4 0.0026 0.0013 3.0E−4 0.0032 0.0012 Lower limit of 95% 1.41 1.36 1.38 1.45 1.32 1.49 1.55 1.30 1.46 CI Upper limit of 95% CI 3.99 3.85 4.94 4.09 3.71 5.16 4.41 3.65 4.71 OR Quartile 3 1.99 1.69 2.37 2.08 1.69 2.49 2.08 1.74 2.44 p Value 0.0028 0.021 5.8E−4 0.0018 0.021 2.0E−4 0.0022 0.015 1.8E−4 Lower limit of 95% 1.27 1.08 1.45 1.31 1.08 1.54 1.30 1.11 1.53 CI Upper limit of 95% CI 3.14 2.65 3.88 3.29 2.65 4.03 3.32 2.74 3.90 OR Quartile 4 3.03 2.83 3.31 2.84 2.74 3.14 2.82 3.08 2.99 p Value 1.6E−4 2.6E−4 1.0E−5 4.7E−4 4.1E−4 2.1E−5 7.9E−4 1.3E−4 4.2E−5 Lower limit of 95% 1.70 1.62 1.95 1.58 1.57 1.86 1.54 1.73 1.77 CI Upper limit of 95% CI 5.38 4.96 5.63 5.08 4.79 5.32 5.17 5.47 5.06

TABLE 10.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11900 19800 11900 20000 11900 20000 Average 20700 42300 20600 42000 20600 42000 Stdev 38800 67400 39100 66700 39100 66700 p (t-test) 3.4E−4 3.9E−4 3.9E−4 Min 200 1490 200 1490 200 1490 Max 465000 428000 465000 428000 465000 428000 n (Patient) 188 129 185 132 185 132 sCr only Median 11800 18800 12000 18300 12000 18500 Average 18600 40600 18700 40000 18500 40200 Stdev 22400 70800 22500 70200 22300 70000 p (t-test) 2.0E−4 3.4E−4 2.6E−4 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 160 157 157 160 156 161 UO only Median 11500 21500 11500 21500 11400 21500 Average 20500 48000 20500 47400 20000 47400 Stdev 37700 72900 37900 72000 37500 71500 p (t-test) 1.4E−5 1.8E−5 1.1E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 211 104 208 107 205 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.62 0.68 0.64 0.61 0.68 0.64 0.62 0.68 SE 0.032 0.031 0.033 0.032 0.031 0.033 0.032 0.031 0.033 p Value 3.2E−5 1.8E−4 9.2E−8 1.6E−5 4.8E−4 5.0E−8 1.6E−5 2.4E−4 2.9E−8 nCohort Recovered 188 160 211 185 157 208 185 156 205 nCohort Non- 129 157 104 132 160 107 132 161 110 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 83% 82% 85% 83% 81% 85% 83% 81% 85% Specificity 30% 31% 30% 31% 31% 30% 31% 31% 30% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 60% 57% 65% 60% 56% 65% 60% 56% 65% Specificity 56% 56% 57% 57% 55% 58% 57% 56% 58% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 36% 34% 41% 36% 34% 41% 36% 34% 41% Specificity 82% 84% 83% 82% 83% 83% 82% 84% 83% OR Quartile 2 2.12 2.01 2.34 2.23 1.97 2.47 2.23 2.00 2.37 p Value 0.0080 0.0092 0.0062 0.0046 0.011 0.0035 0.0046 0.0092 0.0046 Lower limit of 95% 1.22 1.19 1.27 1.28 1.17 1.35 1.28 1.19 1.31 CI Upper limit of 95% CI 3.68 3.39 4.30 3.87 3.31 4.54 3.87 3.37 4.31 OR Quartile 3 1.91 1.68 2.54 1.96 1.56 2.58 1.96 1.60 2.62 p Value 0.0051 0.022 1.8E−4 0.0037 0.050 1.2E−4 0.0037 0.038 8.6E−5 Lower limit of 95% 1.21 1.08 1.56 1.24 1.00 1.59 1.24 1.03 1.62 CI Upper limit of 95% CI 3.02 2.62 4.13 3.08 2.43 4.19 3.08 2.49 4.24 OR Quartile 4 2.51 2.70 3.43 2.55 2.57 3.45 2.55 2.72 3.48 p Value 4.9E−4 2.6E−4 5.3E−6 4.1E−4 5.3E−4 4.5E−6 4.1E−4 2.7E−4 3.8E−6 Lower limit of 95% 1.50 1.58 2.02 1.52 1.51 2.03 1.52 1.59 2.05 CI Upper limit of 95% CI 4.21 4.61 5.82 4.28 4.38 5.86 4.28 4.65 5.91

TABLE 10.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered”cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 94400 115000 94100 114000 92400 114000 Average 101000 126000 101000 125000 99500 125000 Stdev 32200 45000 31900 44800 32900 44600 p (t-test) 2.7E−4 0.0010 8.8E−4 Min 57600 57000 57600 57000 57600 57000 Max 213000 299000 213000 299000 213000 299000 n (Patient) 52 275 42 285 38 289 sCr only Median 96000 116000 96000 116000 96000 115000 Average 101000 127000 101000 127000 101000 127000 Stdev 26600 46300 26000 46300 26400 46100 p (t-test) 1.3E−5 1.1E−5 2.6E−5 Min 57600 57000 57600 57000 57600 57000 Max 213000 299000 213000 299000 213000 299000 n (Patient) 69 257 67 259 63 263 UO only Median 104000 121000 104000 120000 102000 119000 Average 117000 130000 115000 131000 115000 129000 Stdev 43200 44400 41100 46100 41900 45300 p (t-test) 0.0058 9.9E−4 0.0047 Min 57000 64800 57000 64800 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 197 128 176 149 159 166 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.68 0.61 0.70 0.69 0.62 0.72 0.69 0.61 SE 0.035 0.033 0.032 0.039 0.033 0.031 0.038 0.034 0.031 p Value 2.0E−8 2.3E−8 3.9E−4 4.2E−7 1.6E−8 1.4E−4 1.6E−8 4.7E−8 3.3E−4 nCohort Recovered 52 69 197 42 67 176 38 63 159 nCohort Non- 275 257 128 285 259 149 289 263 166 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 78% 78% 85% 78% 78% 84% 78% 78% 83% Specificity 42% 38% 31% 43% 37% 32% 47% 38% 33% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 55% 56% 60% 54% 56% 60% 54% 55% 58% Specificity 77% 71% 56% 76% 72% 58% 79% 71% 58% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 28% 30% 32% 28% 30% 32% 27% 30% 30% Specificity 92% 93% 79% 93% 94% 80% 92% 94% 79% OR Quartile 2 2.63 2.17 2.63 2.59 2.11 2.49 3.16 2.18 2.46 p Value 0.0023 0.0077 9.1E−4 0.0055 0.011 8.9E−4 0.0012 0.0094 7.3E−4 Lower limit of 95% 1.41 1.23 1.49 1.32 1.19 1.46 1.58 1.21 1.46 CI Upper limit of 95% 4.89 3.84 4.67 5.07 3.75 4.28 6.34 3.91 4.16 CI OR Quartile 3 4.12 3.07 1.95 3.76 3.16 2.04 4.40 3.07 1.98 p Value 5.4E−5 1.3E−4 0.0038 5.1E−4 1.1E−4 0.0016 3.6E−4 2.4E−4 0.0024 Lower limit of 95% 2.07 1.73 1.24 1.78 1.76 1.31 1.95 1.69 1.27 CI Upper limit of 95% 8.19 5.46 3.06 7.94 5.68 3.19 9.92 5.59 3.08 CI OR Quartile 4 4.75 5.48 1.79 4.99 6.79 1.86 4.39 6.22 1.60 p Value 0.0037 4.4E−4 0.024 0.0088 3.3E−4 0.017 0.016 6.2E−4 0.070 Lower limit of 95% 1.66 2.12 1.08 1.50 2.39 1.12 1.31 2.18 0.962 CI Upper limit of 95% 13.6 14.1 2.97 16.6 19.3 3.08 14.7 17.7 2.66 CI

TABLE 10.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where revovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 97900 116000 97400 115000 96000 114000 Average 108000 127000 110000 125000 110000 125000 Stdev 35600 45800 37400 45400 38900 44900 p (t-test) 4.8E−4 0.0059 0.012 Min 57600 57000 57600 57000 57600 57000 Max 257000 299000 257000 299000 257000 299000 n (Patient) 87 240 76 251 69 258 sCr only Median 98100 117000 98100 117000 98000 117000 Average 107000 129000 106000 128000 107000 128000 Stdev 32600 46900 32400 46800 32900 46600 p (t-test) 2.4E−5 2.4E−5 8.9E−5 Min 57600 57000 57600 57000 57600 57000 Max 257000 299000 257000 299000 257000 299000 n (Patient) 101 226 99 228 94 233 UO only Median 105000 123000 105000 120000 104000 120000 Average 117000 133000 116000 131000 116000 129000 Stdev 42700 45300 42100 45900 42900 44600 p (t-test) 0.0019 0.0029 0.0066 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 217 108 198 127 181 144 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.66 0.62 0.62 0.66 0.61 0.63 0.65 0.61 SE 0.033 0.031 0.034 0.035 0.031 0.032 0.036 0.032 0.032 p Value 5.8E−6 4.8E−7 2.1E−4 3.4E−4 4.5E−7 5.1E−4 4.5E−4 3.5E−6 3.6E−4 nCohort Recovered 87 101 217 76 99 198 69 94 181 nCohort Non- 240 226 108 251 228 127 258 233 144 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 78% 79% 86% 78% 79% 84% 78% 79% 84% Specificity 34% 35% 30% 34% 34% 31% 36% 34% 32% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 57% 58% 62% 55% 57% 60% 54% 57% 59% Specificity 69% 66% 56% 66% 67% 56% 65% 66% 57% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 29% 30% 34% 27% 30% 32% 27% 30% 31% Specificity 85% 86% 79% 83% 87% 79% 81% 86% 79% OR Quartile 2 1.90 2.02 2.71 1.81 1.96 2.38 2.00 1.89 2.48 p Value 0.019 0.0082 0.0015 0.037 0.012 0.0026 0.017 0.018 0.0011 Lower limit of 95% 1.11 1.20 1.46 1.03 1.16 1.35 1.13 1.11 1.44 CI Upper limit of 95% 3.26 3.40 5.02 3.17 3.31 4.19 3.55 3.21 4.28 CI OR Quartile 3 2.96 2.67 2.06 2.35 2.70 1.90 2.22 2.53 1.90 p Value 4.6E−5 8.6E−5 0.0027 0.0018 7.9E−5 0.0054 0.0046 2.6E−4 0.0045 Lower limit of 95% 1.76 1.64 1.28 1.38 1.65 1.21 1.28 1.54 1.22 CI Upper limit of 95% 4.98 4.35 3.30 4.01 4.42 2.99 3.87 4.17 2.96 CI OR Quartile 4 2.30 2.67 1.99 1.84 2.87 1.83 1.57 2.62 1.66 p Value 0.012 0.0023 0.0088 0.070 0.0014 0.020 0.18 0.0036 0.050 Lower limit of 95% 1.20 1.42 1.19 0.951 1.50 1.10 0.810 1.37 1.00 CI Upper limit of 95% 4.41 5.03 3.33 3.55 5.49 3.03 3.05 5.02 2.74 CI

TABLE 10.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 98600 117000 98600 117000 98000 116000 Average 112000 127000 112000 126000 112000 126000 Stdev 40700 45000 40800 44900 41900 44400 p (t-test) 0.0049 0.0082 0.012 Min 57600 57000 57600 57000 57600 57000 Max 257000 299000 257000 299000 257000 299000 n (Patient) 111 216 104 223 94 233 sCr only Median 98600 118000 98600 118000 98600 118000 Average 111000 129000 110000 129000 109000 129000 Stdev 38500 45900 37700 46000 37600 46000 p (t-test) 2.8E−4 1.2E−4 1.6E−4 Min 57600 57000 57600 57000 57600 57000 Max 257000 299000 257000 299000 257000 299000 n (Patient) 124 203 121 206 116 211 UO only Median 104000 125000 103000 125000 101000 123000 Average 115000 137000 114000 136000 114000 134000 Stdev 41400 46000 40600 46800 41200 45800 p (t-test) 1.9E−5 1.7E−5 7.8E−5 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 219 106 208 117 193 132 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.62 0.64 0.67 0.62 0.65 0.66 0.62 0.64 0.65 SE 0.032 0.031 0.033 0.032 0.031 0.032 0.033 0.031 0.031 p Value 9.1E−5 5.0E−6 5.3E−7 2.7E−4 1.7E−6 5.8E−7 2.2E−4 3.4E−6 8.7E−7 nCohort Recovered 111 124 219 104 121 208 94 116 193 nCohort Non- 216 203 106 223 206 117 233 211 132 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 78% 79% 89% 78% 79% 87% 78% 78% 86% Specificity 32% 31% 32% 31% 31% 32% 33% 31% 33% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 58% 59% 66% 57% 59% 65% 57% 59% 64% Specificity 66% 65% 58% 65% 65% 58% 66% 66% 59% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 29% 31% 39% 29% 31% 37% 28% 31% 34% Specificity 83% 84% 81% 83% 85% 81% 82% 85% 81% OR Quartile 2 1.66 1.71 3.60 1.54 1.69 3.16 1.76 1.61 3.07 p Value 0.055 0.039 1.6E−4 0.11 0.044 2.5E−4 0.037 0.066 1.6E−4 Lower limit of 95% 0.990 1.03 1.85 0.912 1.01 1.71 1.03 0.968 1.72 CI Upper limit of 95% 2.77 2.83 7.01 2.59 2.80 5.85 2.98 2.69 5.49 CI OR Quartile 3 2.69 2.63 2.63 2.55 2.73 2.58 2.53 2.71 2.53 p Value 4.7E−5 4.2E−5 8.5E−5 1.5E−4 2.4E−5 7.6E−5 2.6E−4 3.4E−5 6.9E−5 Lower limit of 95% 1.67 1.66 1.63 1.57 1.71 1.61 1.54 1.69 1.60 CI Upper limit of 95% 4.33 4.17 4.27 4.13 4.35 4.12 4.17 4.34 3.99 CI OR Quartile 4 1.99 2.29 2.74 1.92 2.58 2.52 1.75 2.59 2.18 p Value 0.019 0.0041 1.4E−4 0.028 0.0014 4.1E−4 0.066 0.0016 0.0026 Lower limit of 95% 1.12 1.30 1.63 1.07 1.44 1.51 0.964 1.43 1.31 CI Upper limit of 95% 3.54 4.02 4.60 3.45 4.61 4.20 3.19 4.69 3.62 CI

TABLE 10.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum-creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 98600 119000 98600 118000 98100 118000 Average 112000 128000 112000 128000 111000 128000 Stdev 40900 45100 41200 44900 39400 45300 p (t-test) 0.0012 0.0023 8.2E−4 Min 57600 57000 57600 57000 57600 57000 Max 269000 299000 269000 299000 257000 299000 n (Patient) 134 193 125 202 113 214 sCr only Median 98700 120000 98600 121000 98700 120000 Average 111000 130000 110000 130000 110000 130000 Stdev 39400 45800 38700 45900 38400 46000 p (t-test) 1.2E−4 3.3E−5 5.7E−5 Min 57600 57000 57600 57000 57600 57000 Max 269000 299000 269000 299000 269000 299000 n (Patient) 139 188 137 190 133 194 UO only Median 103000 128000 103000 124000 103000 123000 Average 114000 139000 114000 136000 114000 134000 Stdev 39800 47800 40400 47200 41200 46000 p (t-test) 9.4E−7 1.2E−5 7.2E−5 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 220 105 211 114 198 127 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.65 0.68 0.63 0.66 0.66 0.64 0.65 0.65 SE 0.031 0.030 0.033 0.031 0.030 0.032 0.031 0.030 0.032 p Value 8.4E−6 9.0E−7 4.4E−8 2.0E−5 1.2E−7 5.9E−7 5.3E−6 5.9E−7 1.1E−6 nCohort Recovered 134 139 220 125 137 211 113 133 198 nCohort Non- 193 188 105 202 190 114 214 194 127 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 80% 80% 89% 80% 81% 87% 80% 79% 86% Specificity 33% 32% 31% 33% 33% 31% 35% 32% 32% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 61% 62% 67% 60% 62% 64% 59% 61% 64% Specificity 66% 65% 58% 66% 66% 57% 66% 66% 59% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 31% 32% 40% 30% 33% 38% 30% 32% 35% Specificity 83% 84% 82% 83% 85% 82% 84% 86% 81% OR Quartile 2 1.99 1.95 3.54 1.92 2.02 3.00 2.10 1.78 2.83 p Value 0.0075 0.0094 1.9E−4 0.012 0.0064 4.7E−4 0.0046 0.026 4.6E−4 Lower limit of 95% 1.20 1.18 1.82 1.15 1.22 1.62 1.26 1.07 1.58 CI Upper limit of 95% 3.31 3.24 6.89 3.18 3.35 5.56 3.50 2.94 5.05 CI OR Quartile 3 3.01 3.05 2.73 2.85 3.24 2.39 2.83 3.10 2.49 p Value 2.6E−6 1.7E−6 5.1E−5 9.8E−6 5.4E−7 2.7E−4 1.9E−5 1.5E−6 9.8E−5 Lower limit of 95% 1.90 1.93 1.68 1.79 2.05 1.50 1.76 1.96 1.57 CI Upper limit of 95% 4.76 4.82 4.44 4.53 5.14 3.83 4.55 4.92 3.94 CI OR Quartile 4 2.12 2.49 3.00 2.14 2.83 2.67 2.25 2.89 2.23 p Value 0.0066 0.0011 3.4E−5 0.0073 2.9E−4 1.8E−4 0.0063 2.8E−4 0.0020 Lower limit of 95% 1.23 1.44 1.78 1.23 1.61 1.60 1.26 1.63 1.34 CI Upper limit of 95% 3.66 4.32 5.04 3.74 4.98 4.47 4.03 5.11 3.71 CI

TABLE 10.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 101000 119000 101000 120000 101000 119000 Average 114000 132000 114000 132000 114000 131000 Stdev 40800 46600 40900 46200 40900 46200 p (t-test) 4.5E−4 3.1E−4 4.3E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 187 140 184 143 183 144 sCr only Median 101000 118000 99700 120000 100000 119000 Average 113000 130000 112000 131000 112000 130000 Stdev 38800 47200 37800 47400 37900 47400 p (t-test) 4.7E−4 6.7E−5 1.5E−4 Min 57600 57000 57600 57000 57600 57000 Max 269000 299000 269000 299000 269000 299000 n (Patient) 159 168 156 171 154 173 UO only Median 103000 124000 103000 125000 103000 124000 Average 114000 138000 114000 137000 114000 137000 Stdev 39600 48200 39800 47700 40100 47300 p (t-test) 3.0E−6 3.1E−6 5.1E−6 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 215 110 212 113 209 116 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.62 0.66 0.63 0.64 0.67 0.63 0.63 0.67 SE 0.031 0.031 0.033 0.031 0.030 0.032 0.031 0.031 0.032 p Value 5.0E−5 8.3E−5 4.7E−7 1.9E−5 8.0E−6 2.1E−7 3.2E−5 3.2E−5 2.0E−7 nCohort 187 159 215 184 156 212 183 154 209 Recovered nCohort Non- 140 168 110 143 171 113 144 173 116 recovered Cutoff 90200 90200 90200 90200 90200 90200 90200 90200 90200 Quartile 2 Sensitivity 83% 80% 86% 83% 81% 87% 83% 80% 87% Specificity 31% 31% 31% 32% 31% 31% 31% 31% 32% Cutoff 109000 109000 109000 109000 109000 109000 109000 109000 109000 Quartile 3 Sensitivity 61% 60% 65% 61% 60% 65% 60% 60% 66% Specificity 58% 60% 58% 58% 61% 58% 58% 60% 58% Cutoff 137000 137000 137000 137000 137000 137000 137000 137000 137000 Quartile 4 Sensitivity 33% 32% 39% 34% 33% 39% 33% 33% 38% Specificity 81% 82% 82% 82% 84% 82% 81% 84% 82% OR Quartile 2 2.17 1.82 2.81 2.28 1.92 2.95 2.15 1.73 3.11 p Value 0.0047 0.021 0.0010 0.0026 0.012 5.7E−4 0.0048 0.033 3.1E−4 Lower limit of 1.27 1.10 1.51 1.33 1.15 1.59 1.26 1.04 1.68 95% CI Upper limit of 3.72 3.03 5.20 3.91 3.18 5.47 3.67 2.87 5.75 95% CI OR Quartile 3 2.11 2.18 2.58 2.16 2.36 2.62 2.10 2.24 2.66 p Value 0.0010 5.4E−4 9.8E−5 7.1E−4 1.5E−4 6.7E−5 0.0011 3.5E−4 4.6E−5 Lower limit of 1.35 1.40 1.60 1.38 1.51 1.63 1.35 1.44 1.66 95% CI Upper limit of 3.30 3.40 4.16 3.37 3.68 4.21 3.28 3.49 4.27 95% CI OR Quartile 4 2.05 2.22 2.90 2.23 2.62 2.92 2.19 2.54 2.75 p Value 0.0054 0.0028 5.5E−5 0.0020 4.0E−4 4.7E−5 0.0025 6.2E−4 1.1E−4 Lower limit of 1.24 1.32 1.73 1.34 1.54 1.74 1.32 1.49 1.64 95% CI Upper limit of 3.41 3.73 4.86 3.71 4.46 4.89 3.64 4.32 4.60 95% CI

Example 11. Use of Metalloproteinase Inhibitor 2 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 2 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 11.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 10600 17900 10400 17900 10100 17900 Average 15300 36900 14700 36700 14500 36400 Stdev 13900 63700 13300 63200 13300 62700 p (t-test) 5.8E−4 5.1E−4 6.3E−4 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 108 209 104 213 100 217 sCr only Median 12000 16000 11800 16000 11500 16600 Average 17000 37300 17000 37100 17000 36900 Stdev 14700 66400 14800 66100 14900 65800 p (t-test) 8.2E−4 9.5E−4 0.0011 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 126 190 124 192 122 194 UO only Median 11400 25100 11000 22700 11000 21700 Average 20500 51500 19900 48500 19900 46000 Stdev 38100 74800 38900 70600 39600 68200 p (t-test) 1.9E−6 5.3E−6 2.4E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 222 93 208 107 198 117 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.59 0.74 0.66 0.60 0.74 0.66 0.60 0.72 SE 0.031 0.032 0.033 0.031 0.032 0.031 0.031 0.032 0.031 p Value 1.6E−6 0.0031 5.1E−13 2.3E−7 0.0023 5.3E−15 1.3E−7 0.0017 2.8E−13 nCohort Recovered 108 126 222 104 124 208 100 122 198 nCohort Non- 209 190 93 213 192 107 217 194 117 recovered Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 81% 79% 92% 82% 79% 93% 82% 79% 91% Specificity 37% 31% 32% 38% 31% 35% 39% 32% 34% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 56% 53% 71% 56% 54% 69% 57% 54% 68% Specificity 61% 55% 59% 62% 56% 60% 64% 57% 60% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 32% 32% 44% 32% 32% 43% 32% 31% 41% Specificity 87% 86% 83% 88% 85% 84% 89% 85% 84% OR Quartile 2 2.56 1.68 5.90 2.79 1.74 7.56 2.83 1.81 5.04 p Value 4.2E−4 0.048 2.2E−5  1.3E−4 0.034 1.2E−6  1.1E−4 0.024 3.9E−6  Lower limit of 95% 1.52 1.01 2.60 1.65 1.04 3.34 1.67 1.08 2.54 CI Upper limit of 4.33 2.81 13.4 4.72 2.92 17.1 4.80 3.03 10.0 95% CI OR Quartile 3 2.00 1.37 3.45 2.15 1.45 3.31 2.33 1.54 3.13 p Value 0.0042 0.17 3.2E−6  0.0018 0.11 2.1E−6  7.1E−4 0.065 3.2E−6  Lower limit of 95% 1.24 0.874 2.05 1.33 0.922 2.02 1.43 0.974 1.94 CI Upper limit of 3.21 2.16 5.82 3.48 2.29 5.43 3.79 2.42 5.06 95% CI OR Quartile 4 3.10 2.84 3.82 3.60 2.74 4.00 3.77 2.65 3.75 p Value 4.6E−4 4.7E−4 1.1E−6  1.7E−4 7.2E−4 3.6E−7  1.6E−4 0.0011 1.1E−6  Lower limit of 95% 1.65 1.58 2.23 1.85 1.53 2.35 1.89 1.48 2.20 CI Upper limit of 5.83 5.09 6.54 7.01 4.92 6.82 7.51 4.76 6.38 95% CI

TABLE 11.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11300 19300 11100 19300 11000 19300 Average 15500 40000 15400 39800 14900 39500 Stdev 13800 67800 13800 67400 13300 66600 p (t-test) 4.4E−5 4.7E−5 4.7E−5 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 136 181 134 183 129 188 sCr only Median 12000 18700 12000 18600 11800 18200 Average 16900 40600 16900 40500 16900 40200 Stdev 14700 70200 14700 70000 14800 69600 p (t-test) 6.5E−5 7.4E−5 8.9E−5 Min 200 658 200 658 200 658 Max 76000 465000 76000 465000 76000 465000 n (Patient) 149 168 148 169 146 171 UO only Median 11600 22100 11400 22100 11300 21700 Average 20600 52500 20300 51100 20100 47800 Stdev 37600 76500 38000 74400 38800 70600 p (t-test) 1.3E−6 1.9E−6 9.6E−6 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 226 89 220 95 207 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.61 0.72 0.66 0.61 0.72 0.67 0.62 0.71 SE 0.030 0.031 0.034 0.030 0.031 0.033 0.030 0.031 0.032 p Value 4.5E−7 3.4E−4 2.3E−10 1.8E−7 3.2E−4 2.0E−11 3.8E−8 2.4E−4 3.5E−11 nCohort Recovered 136 149 226 134 148 220 129 146 207 nCohort Non- 181 168 89 183 169 95 188 171 108 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 81% 79% 89% 81% 79% 89% 81% 80% 88% Specificity 33% 30% 31% 34% 30% 31% 34% 30% 32% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 59% 56% 69% 59% 56% 68% 60% 57% 68% Specificity 61% 56% 57% 62% 57% 58% 64% 58% 59% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 34% 35% 44% 34% 34% 44% 35% 34% 43% Specificity 87% 85% 82% 87% 85% 83% 88% 85% 84% OR Quartile 2 2.14 1.59 3.47 2.22 1.62 3.88 2.26 1.68 3.42 p Value 0.0039 0.075 6.6E−4  0.0026 0.065 2.0E−4  0.0020 0.048 2.0E−4  Lower limit of 95% 1.28 0.954 1.70 1.32 0.970 1.90 1.35 1.00 1.79 CI Upper limit of 95% CI 3.58 2.66 7.11 3.72 2.70 7.94 3.80 2.80 6.55 OR Quartile 3 2.21 1.64 2.90 2.34 1.68 2.96 2.57 1.78 2.99 p Value 6.1E−4 0.029 5.9E−5  2.6E−4 0.022 2.9E−5  6.2E−5 0.012 1.1E−5  Lower limit of 95% 1.41 1.05 1.72 1.48 1.08 1.78 1.62 1.14 1.84 CI Upper limit of 95% CI 3.49 2.56 4.87 3.70 2.63 4.92 4.08 2.78 4.88 OR Quartile 4 3.42 3.04 3.63 3.61 2.99 3.92 4.02 2.89 3.91 p Value 3.7E−5 8.0E−5 3.0E−6  2.2E−5 1.0E−4 6.3E−7  9.9E−6 1.7E−4 5.2E−7  Lower limit of 95% 1.91 1.75 2.11 2.00 1.72 2.29 2.17 1.66 2.30 CI Upper limit of 95% CI 6.12 5.29 6.23 6.54 5.20 6.71 7.44 5.03 6.67

TABLE 11.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11300 19300 11300 19300 11000 19300 Average 17200 41100 17300 40700 16800 40600 Stdev 21500 69500 21700 69200 21600 68400 p (t-test) 5.7E−5 7.8E−5 6.3E−5 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 154 163 152 165 148 169 sCr only Median 11500 19000 11500 18900 11400 18800 Average 17900 41500 17900 41300 17900 41200 Stdev 21500 71000 21600 70800 21600 70600 p (t-test) 7.0E−5 8.0E−5 8.9E−5 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 161 156 160 157 159 158 UO only Median 11900 21500 11900 21500 11500 21500 Average 21500 49500 21600 49100 21200 46900 Stdev 38600 75700 38600 75400 39400 71800 p (t-test) 2.1E−5 2.7E−5 5.4E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 224 91 223 92 212 103 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.63 0.68 0.65 0.63 0.68 0.67 0.63 0.69 SE 0.0030 0.031 0.035 0.030 0.031 0.035 0.030 0.031 0.033 p Value 3.0E−7 2.0E−5 2.2E−7 4.5E−7 1.8E−7 3.3E−7 3.5E−8 1.6E−5 1.3E−8 nCohort Recovered 154 161 224 152 160 223 148 159 212 nCohort Non- 163 156 91 165 157 92 169 158 103 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 83% 81% 86% 83% 82% 86% 83% 82% 87% Specificity 33% 31% 29% 34% 31% 30% 34% 31% 31% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 60% 58% 66% 60% 59% 65% 61% 59% 66% Specificity 60% 58% 56% 61% 58% 56% 62% 58% 58% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 36% 37% 42% 36% 36% 41% 36% 36% 41% Specificity 86% 86% 82% 86% 86% 82% 87% 86% 83% OR Quartile 2 2.39 1.97 2.51 2.47 2.01 2.55 2.65 2.04 3.13 p Value 0.0012 0.011 0.0059 7.8E−4 0.0092 0.0049 3.1E−4 0.0076 5.8E−4 Lower limit of 95% 1.41 1.17 1.30 1.46 1.19 1.33 1.56 1.21 1.63 CI Upper limit of 95% CI 4.05 3.33 4.82 4.19 3.39 4.91 4.49 3.45 6.00 OR Quartile 3 2.30 1.91 2.49 2.30 1.96 2.39 2.56 2.02 2.63 p Value 2.9E−4 0.0043 4.3E−4 2.9E−4 0.0030 7.0E−4 4.8E−5 0.0021 1.1E−4 Lower limit of 95% 1.47 1.23 1.50 1.47 1.26 1.44 1.63 12.9 1.61 CI Upper limit of 95% CI 3.61 2.99 4.13 3.61 3.07 3.96 4.04 3.15 4.30 OR Quartile 4 3.59 3.45 3.20 3.47 3.40 3.12 3.83 3.34 3.26 p Value 7.7E−6 9.5E−6 2.2E−5 1.3E−5 1.3E−5 3.1E−5 4.6E−6 1.7E−5 1.2E−5 Lower limit of 95% 2.05 2.00 1.87 1.98 1.96 1.83 2.16 1.93 1.92 CI Upper limit of 95% CI 6.29 5.98 5.47 6.08 5.88 5.34 6.81 5.77 5.53

TABLE 11.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11500 19100 11400 19000 11300 19200 Average 19000 41300 18900 40900 18600 40800 Stdev 25200 71400 25300 70700 25400 69900 p (t-test) 1.8E−4 2.2E−4 1.8E−4 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 168 149 165 152 161 156 sCr only Median 11900 18800 11900 18700 11800 18600 Average 19300 41600 19300 41400 19300 41200 Stdev 25100 72300 25100 72100 25200 71900 p (t-test) 1.9E−4 2.2E−4 2.4E−4 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 172 145 171 146 170 147 UO only Median 11900 21500 11800 21500 11500 21500 Average 21600 49100 21500 49100 21300 47200 Stdev 38600 75500 38600 75100 39200 72500 p (t-test) 2.6E−5 2.4E−5 5.3E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 223 92 222 93 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.62 0.67 0.64 0.62 0.68 0.65 0.62 0.68 SE 0.031 0.032 0.035 0.031 0.032 0.035 0.031 0.032 0.033 p Value 1.0E−5 1.2E−4 6.6E−7 7.2E−6 1.1E−4 2.8E−7 8.9E−7 1.0E−4 5.3E−8 nCohort Recovered 168 172 223 165 171 222 161 170 214 nCohort Non- 149 145 92 152 146 93 156 147 101 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 83% 81% 85% 83% 82% 85% 83% 82% 86% Specificity 32% 30% 29% 32% 30% 29% 33% 31% 30% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 60% 58% 65% 60% 58% 66% 61% 59% 66% Specificity 58% 56% 56% 59% 57% 56% 60% 57% 57% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 36% 37% 41% 36% 36% 42% 36% 36% 41% Specificity 84% 84% 82% 84% 84% 82% 85% 84% 82% OR Quartile 2 2.18 1.89 2.29 2.29 1.93 2.34 2.45 1.96 2.71 p Value 0.0042 0.018 0.011 0.0023 0.015 0.0091 9.9E−4 0.013 0.0021 Lower limit of 95% CI 1.28 1.11 1.21 1.34 1.13 1.23 1.44 1.15 1.44 Upper limit of 95% CI 3.72 3.22 4.34 3.91 3.27 4.42 4.19 3.33 5.12 OR Quartile 3 2.08 1.78 2.39 2.13 1.83 2.46 2.36 1.87 2.66 p Value 0.0014 0.011 7.0E−4 9.7E−4 0.0082 4.7E−4 1.9E−4 0.0059 1.0E−4 Lower limit of 95% CI 1.33 1.14 1.44 1.36 1.17 1.48 1.50 1.20 1.63 Upper limit of 95% CI 3.25 2.78 3.96 3.33 2.86 4.06 3.70 2.93 4.36 OR Quartile 4 2.88 3.09 3.12 2.95 3.04 3.29 3.20 2.99 3.16 p Value 9.3E−5 3.2E−5 3.1E−5 7.4E−5 4.2E−5 1.3E−5 2.8E−5 5.5E−5 2.0E−5 Lower limit of 95% CI 1.70 1.82 1.83 1.73 1.79 1.92 1.86 1.76 1.86 Upper limit of 95% CI 4.90 5.27 5.34 5.03 5.17 5.61 5.50 5.08 5.38

TABLE 11.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11900 20900 11900 20900 11800 21100 Average 21500 43700 21400 43400 21400 43300 Stdev 39800 69300 39900 68800 40000 68500 p (t-test) 3.4E−4 3.7E−4 4.0E−4 Min 200 1490 200 1490 200 1490 Max 465000 428000 465000 428000 465000 428000 n (Patient) 203 114 201 116 200 117 sCr only Median 11900 19000 11900 19000 11900 19000 Average 19100 44000 19100 44000 19100 44000 Stdev 24400 75300 24400 75300 24400 75300 p (t-test) 3.4E−5 3.4E−5 3.4E−5 Min 200 658 200 658 200 658 Max 215000 465000 215000 465000 215000 465000 n (Patient) 185 132 185 132 185 132 UO only Median 11800 21500 11700 21500 11700 21500 Average 21700 47200 21600 47200 21600 46600 Stdev 39100 73500 39200 73100 39400 72500 p (t-test) 7.4E−5 6.6E−5 9.6E−5 Min 200 2170 200 2170 200 2170 Max 465000 428000 465000 428000 465000 428000 n (Patient) 217 98 216 99 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.63 0.67 0.64 0.63 0.67 0.65 0.63 0.67 SE 0.033 0.032 0.034 0.033 0.032 0.034 0.033 0.032 0.034 p Value 1.4E−5 4.9E−5 1.2E−6 1.1E−5 4.9E−5 5.1E−7 8.0E−6 4.9E−5 6.9E−7 nCohort Recovered 203 185 217 201 185 216 200 185 214 nCohort Non- 114 132 98 116 132 99 117 132 101 recovered Cutoff Quartile 2 7990 7990 8020 7990 7990 8020 7990 7990 8020 Sensitivity 83% 83% 84% 84% 83% 84% 84% 83% 84% Specificity 30% 30% 29% 30% 30% 29% 30% 30% 29% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 62% 59% 65% 62% 59% 66% 62% 59% 65% Specificity 57% 56% 57% 57% 56% 57% 57% 56% 57% Cutoff Quartile 4 28900 28900 29400 28900 28900 29400 28900 28900 29400 Sensitivity 39% 39% 41% 39% 39% 41% 38% 39% 41% Specificity 82% 84% 82% 83% 84% 82% 82% 84% 82% OR Quartile 2 2.10 2.06 2.10 2.17 2.06 2.14 2.21 2.06 2.22 p Value 0.012 0.0099 0.017 0.0084 0.0099 0.015 0.0070 0.0099 0.011 Lower limit of 95% CI 1.18 1.19 1.14 1.22 1.19 1.16 1.24 1.19 1.20 Upper limit of 95% CI 3.74 3.56 3.86 3.87 3.56 3.93 3.94 3.56 4.08 OR Quartile 3 2.16 1.85 2.46 2.14 1.85 2.53 2.20 1.85 2.50 p Value 0.0013 0.0075 3.6E−4 0.0014 0.0075 2.4E−4 9.5E−4 0.0075 2.5E−4 Lower limit of 95% CI 1.35 1.18 1.50 1.34 1.18 1.54 1.38 1.18 1.53 Upper limit of 95% CI 3.45 2.92 4.04 3.42 2.92 4.15 3.51 2.92 4.09 OR Quartile 4 2.92 3.39 3.15 3.01 3.39 3.31 2.95 3.39 3.16 p Value 5.8E−5 6.2E−6 2.3E−5 3.6E−5 6.2E−6 1.0E−5 4.8E−5 6.2E−6 2.0E−5 Lower limit of 95% CI 1.73 2.00 1.85 1.78 2.00 1.95 1.75 2.00 1.86 Upper limit of 95% CI 4.91 5.75 5.36 5.07 5.75 5.63 4.96 5.75 5.38

TABLE 11.6 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 97900 117000 97500 117000 97500 117000 Average 107000 129000 107000 128000 107000 128000 Stdev 34500 46400 34800 46300 35400 46100 p (t-test) 2.0E−5 4.4E−5 1.0E−4 Min 57600 57000 57600 57000 57600 57000 Max 257000 299000 257000 299000 257000 299000 n (Patient) 105 222 102 225 98 229 sCr only Median 97900 120000 97900 120000 97900 120000 Average 106000 131000 106000 131000 106000 131000 Stdev 32400 47500 32700 47300 32600 47200 p (t-test) 5.5E−7 7.3E−7 6.2E−7 Min 57600 57000 57600 57000 57600 57000 Max 257000 299000 257000 299000 257000 299000 n (Patient) 121 205 119 207 117 209 UO only Median 106000 123000 105000 124000 104000 123000 Average 118000 132000 116000 134000 115000 133000 Stdev 43100 45000 41300 46900 41800 46000 p (t-test) 0.0068 2.9E−4 6.0E−4 Min 57000 70200 57000 70200 57000 70200 Max 299000 281000 299000 281000 299000 281000 n (Patient) 226 99 213 112 202 123 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.68 0.62 0.66 0.68 0.63 0.66 0.68 0.63 SE 0.031 0.030 0.035 0.031 0.030 0.033 0.031 0.030 0.032 p Value 7.3E−8 1.9E−9 9.0E−4 2.6E−7 1.6E−9 4.9E−5 7.9E−7 9.3E−10 5.1E−5 nCohort Recovered 105 121 226 102 119 213 98 117 202 nCohort Non- 222 205 99 225 207 112 229 209 123 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 80% 80% 86% 80% 80% 87% 79% 80% 86% Specificity 35% 34% 30% 35% 34% 31% 35% 34% 32% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 59% 60% 61% 58% 60% 62% 58% 60% 60% Specificity 68% 67% 54% 68% 67% 56% 67% 68% 56% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 32% 33% 35% 31% 33% 37% 31% 33% 34% Specificity 89% 88% 79% 88% 88% 81% 88% 89% 80% OR Quartile 2 2.14 2.05 2.56 2.12 2.13 2.90 2.00 2.07 2.89 p Value 0.0039 0.0057 0.0036 0.0045 0.0037 7.0E−4 0.0093 0.0053 4.4E−4 Lower limit of 95% 1.28 1.23 1.36 1.26 1.28 1.57 1.19 1.24 1.60 CI Upper limit of 95% 3.59 3.41 4.82 3.57 3.54 5.38 3.38 3.44 5.23 CI OR Quartile 3 2.95 3.04 1.84 2.91 3.06 2.03 2.81 3.09 1.92 p Value 1.4E−5 3.7E−6 0.013 2.1E−5 3.5E−6 0.0029 4.7E−5 3.3E−6 0.0051 Lower limit of 95% 1.81 1.90 1.14 1.78 1.91 1.27 1.71 1.92 1.22 CI Upper limit of 95% 4.81 4.86 2.97 4.77 4.92 3.24 4.61 4.98 3.02 CI OR Quartile 4 3.57 3.79 2.08 3.39 3.67 2.42 3.16 3.94 2.10 p Value 1.7E−4 3.2E−5 0.0059 3.2E−4 5.0E−5 7.3E−4 7.2E−4 3.0E−5 0.0042 Lower limit of 95% 1.84 2.02 1.24 1.74 1.96 1.45 1.62 2.07 1.26 CI Upper limit of 95% 6.94 7.11 3.51 6.59 6.88 4.05 6.14 7.51 3.49 CI

TABLE 11.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for ihe “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 98000 120000 97900 119000 98000 118000 Average 111000 129000 111000 129000 112000 128000 Stdev 41500 44400 41600 44300 42200 44200 p (t-test) 2.4E−4 2.5E−4 8.6E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 134 193 133 194 128 199 sCr only Median 98600 121000 98600 121000 98600 121000 Average 110000 131000 110000 131000 110000 131000 Stdev 39300 45700 39400 45600 39500 45400 p (t-test) 1.9E−5 2.1E−5 2.0E−5 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 146 181 145 182 143 184 UO only Median 104000 124000 104000 125000 104000 125000 Average 116000 136000 115000 138000 115000 135000 Stdev 41900 46400 40800 47500 41300 46100 p (t-test) 1.5E−4 1.5E−5 4.3E−5 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 231 94 225 100 210 115 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.66 0.65 0.66 0.66 0.67 0.65 0.66 0.66 SE 0.030 0.030 0.035 0.030 0.030 0.034 0.030 0.030 0.032 p Value 2.0E−7 4.7E−8 1.3E−5 1.7E−7 4.0E−8 1.0E−6 1.6E−6 2.8E−8 9.4E−7 nCohort Recovered 134 146 231 133 145 225 128 143 210 nCohort Non- 193 181 94 194 182 100 199 184 115 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 82% 82% 88% 82% 82% 89% 81% 82% 87% Specificity 35% 34% 30% 35% 34% 31% 34% 34% 31% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 61% 62% 65% 61% 62% 66% 60% 62% 64% Specificity 66% 64% 56% 66% 65% 57% 66% 65% 58% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 31% 33% 38% 31% 32% 39% 30% 33% 37% Specificity 84% 84% 80% 83% 84% 81% 83% 85% 81% OR Quartile 2 2.44 2.27 3.28 2.48 2.30 3.65 2.22 2.23 3.06 p Value 6.1E−4 0.0017 7.2E−4 4.8E−4 0.0013 2.2E−4 0.0021 0.0020 3.8E−4 Lower limit of 95% CI 1.46 1.36 1.65 1.49 1.38 1.84 1.34 1.34 1.65 Upper limit of 95% CI 4.06 3.77 6.53 4.14 3.84 7.26 3.69 3.71 5.66 OR Quartile 3 3.01 2.93 2.34 3.10 3.02 2.56 2.90 3.03 2.45 p Value 2.6E−6 3.1E−6 8.1E−4 1.5E−6 1.8E−6 1.7E−4 6.4E−6 1.8E−6 1.8E−4 Lower limit of 95% CI 1.90 1.87 1.42 1.96 1.92 1.57 1.83 1.92 1.53 Upper limit of 95% CI 4.76 4.61 3.84 4.92 4.75 4.18 4.60 4.77 3.92 OR Quartile 4 2.30 2.59 2.50 2.26 2.54 2.71 2.08 2.66 2.45 p Value 0.0030 6.0E−4 6.6E−4 0.0036 7.5E−4 1.8E−4 0.0091 4.7E−4 6.3E−4 Lower limit of 95% CI 1.33 1.50 1.47 1.30 1.48 1.61 1.20 1.54 1.46 Upper limit of 95% CI 3.98 4.45 4.23 3.91 4.38 4.56 3.60 4.61 4.08

TABLE 11.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for ihe “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 98800 120000 98600 120000 98600 120000 Average 114000 129000 114000 129000 114000 128000 Stdev 42700 44200 43000 43900 43300 43800 p (t-test) 0.0018 0.0017 0.0029 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 154 173 152 175 148 179 sCr only Median 99200 122000 99100 121000 99000 122000 Average 113000 131000 113000 131000 112000 131000 Stdev 41100 45200 41200 45000 41200 45000 p (t-test) 1.7E−4 1.9E−4 1.3E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 161 166 160 167 159 168 UO only Median 104000 124000 104000 125000 103000 127000 Average 115000 138000 115000 139000 114000 138000 Stdev 41100 47300 40900 47000 40200 47200 p (t-test) 2.2E−5 7.1E−6 2.8E−6 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 230 95 228 97 215 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.65 0.66 0.64 0.65 0.67 0.63 0.65 0.68 SE 0.030 0.030 0.034 0.030 0.030 0.034 0.030 0.030 0.033 p Value 1.0E−5 1.1E−6 2.2E−6 5.4E−6 9.6E−7 4.5E−7 1.1E−5 4.2E−7 6.7E−8 nCohort Recovered 154 161 230 152 160 228 148 159 215 nCohort Non- 173 166 95 175 167 97 179 168 110 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 83% 83% 88% 83% 83% 89% 82% 83% 87% Specificity 34% 33% 30% 34% 33% 31% 34% 33% 31% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 62% 63% 66% 62% 63% 67% 62% 63% 67% Specificity 63% 63% 57% 64% 63% 57% 64% 64% 59% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 31% 33% 39% 30% 32% 40% 30% 33% 39% Specificity 81% 83% 80% 81% 82% 81% 81% 83% 82% OR Quartile 2 2.43 2.32 3.34 2.51 2.36 3.46 2.34 2.40 3.10 p Value 7.5E−4 0.0015 5.9E−4 4.7E−4 0.0012 4.0E−4 0.0011 9.5E−4 4.3E−4 Lower limit of 95% CI 1.45 1.38 1.68 1.50 1.40 1.74 1.40 1.43 1.65 Upper limit of 95% CI 4.07 3.89 6.65 4.21 3.96 6.89 3.91 4.02 5.83 OR Quartile 3 2.76 2.82 2.56 2.91 2.90 2.69 2.93 2.98 2.91 p Value 9.2E−6 5.7E−6 2.2E−4 3.3E−6 3.4E−6 9.5E−5 3.2E−6 2.0E−6 1.4E−5 Lower limit of 95% CI 1.76 1.80 1.55 1.86 1.85 1.64 1.86 1.90 1.80 Upper limit of 95% CI 4.32 4.42 4.22 4.57 4.54 4.43 4.60 4.67 4.71 OR Quartile 4 1.90 2.29 2.62 1.84 2.25 2.89 1.85 2.38 2.90 p Value 0.015 0.0018 3.2E−4 0.021 0.0022 7.1E−5 0.020 0.0012 5.5E−5 Lower limit of 95% CI 1.13 1.36 1.55 1.10 1.34 1.71 1.10 1.41 1.73 Upper limit of 95% CI 3.19 3.86 4.44 3.09 3.79 4.89 3.12 4.02 4.86

TABLE 11.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for ihe “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 99100 120000 98700 120000 98700 120000 Average 114000 130000 114000 130000 114000 129000 Stdev 42500 44300 42900 44000 43200 43800 p (t-test) 9.0E−4 0.0010 0.0018 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 166 161 163 164 159 168 sCr only Median 99200 122000 99100 121000 99000 122000 Average 113000 131000 113000 131000 113000 131000 Stdev 41400 44900 41600 44800 41500 44700 p (t-test) 1.2E−4 1.4E−4 9.3E−5 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 169 158 168 159 167 160 UO only Median 103000 130000 103000 129000 103000 128000 Average 114000 141000 114000 141000 114000 139000 Stdev 39500 48200 39500 48100 40000 47400 p (t-test) 1.1E−7 1.6E−7 9.5E−7 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 226 99 225 100 217 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.65 0.70 0.64 0.65 0.69 0.64 0.66 0.68 SE 0.030 0.030 0.033 0.030 0.030 0.033 0.030 0.030 0.033 p Value 4.6E−6 5.4E−7 3.6E−9 3.0E−6 4.8E−7 5.6E−9 6.3E−6 2.1E−7 1.9E−8 nCohort Recovered 166 169 226 163 168 225 159 167 217 nCohort Non- 161 158 99 164 159 100 168 160 108 recovered Cutoff Quartile 2 90200 90200 90200 90200 90200 90200 90200 90200 90200 Sensitivity 83% 83% 89% 83% 83% 89% 82% 83% 87% Specificity 33% 33% 31% 33% 33% 31% 33% 33% 31% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 63% 63% 69% 63% 64% 68% 62% 64% 68% Specificity 62% 62% 58% 63% 62% 58% 63% 63% 59% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 32% 33% 42% 31% 33% 42% 31% 33% 40% Specificity 81% 82% 82% 81% 82% 82% 81% 83% 82% OR Quartile 2 2.29 2.34 3.59 2.41 2.38 3.65 2.24 2.42 3.00 p Value 0.0018 0.0015 2.7E−4 9.6E−4 0.0012 2.2E−4 0.0022 9.7E−4 6.5E−4 Lower limit of 1.36 1.39 1.81 1.43 1.41 1.84 1.34 1.43 1.60 95% CI Upper limit of 3.86 3.96 7.14 4.05 4.02 7.26 3.74 4.09 5.64 95% CI OR Quartile 3 2.75 2.83 3.02 2.82 2.90 2.91 2.82 2.98 2.94 p Value 9.3E−6 5.6E−6 1.4E−5 5.7E−6 3.3E−6 2.5E−5 5.7E−6 2.0E−6 1.3E−5 Lower limit of 1.76 1.81 1.83 1.80 1.85 1.77 1.80 1.90 1.81 95% CI Upper limit of 4.31 4.43 4.99 4.42 4.55 4.78 4.42 4.67 4.78 95% CI OR Quartile 4 2.02 2.27 3.43 1.92 2.24 3.35 1.93 2.36 3.02 p Value 0.0072 0.0018 4.3E−6 0.012 0.0022 6.1E−6 0.012 0.0012 2.9E−5 Lower limit of 1.21 1.36 2.03 1.15 1.34 1.98 1.15 1.40 1.80 95% CI Upper limit of 3.37 3.81 5.79 3.21 3.74 5.66 3.22 3.96 5.07 95% CI

TABLE 11.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for ihe “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 101000 123000 101000 123000 101000 123000 Average 114000 135000 114000 135000 114000 135000 Stdev 40600 46500 40800 46200 40800 46100 p (t-test) 1.8E−5 2.1E−5 1.7E−5 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 204 123 202 125 201 126 sCr only Median 101000 121000 101000 121000 101000 121000 Average 114000 131000 114000 131000 114000 131000 Stdev 41500 45600 41500 45600 41500 45600 p (t-test) 4.5E−4 4.5E−4 4.5E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 184 143 184 143 184 143 UO only Median 103000 125000 103000 125000 102000 126000 Average 114000 139000 114000 139000 114000 139000 Stdev 39400 48400 39500 48400 39600 47900 p (t-test) 6.0E−7 8.8E−7 6.3E−7 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 220 105 219 106 217 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.64 0.68 0.66 0.64 0.68 0.66 0.64 0.68 SE 0.032 0.031 0.033 0.032 0.031 0.033 0.031 0.031 0.033 p Value 3.6E−7 1.4E−5 5.9E−8 2.8E−7 1.4E−5 8.4E−8 1.5E−7 1.4E−5 2.5E−8 nCohort 204 184 220 202 184 219 201 184 217 Recovered nCohort Non- 123 143 105 125 143 106 126 143 108 recovered Cutoff 90200 90200 90200 90200 90200 90200 90200 90200 90200 Quartile 2 Sensitivity 86% 83% 87% 86% 83% 87% 87% 83% 87% Specificity 32% 31% 30% 32% 31% 31% 32% 31% 31% Cutoff 109000 109000 109000 109000 109000 109000 109000 109000 109000 Quartile 3 Sensitivity 65% 62% 68% 65% 62% 67% 65% 62% 68% Specificity 59% 59% 58% 59% 59% 58% 59% 59% 59% Cutoff 137000 137000 137000 137000 137000 137000 137000 137000 137000 Quartile 4 Sensitivity 36% 34% 40% 35% 34% 40% 36% 34% 40% Specificity 81% 82% 82% 81% 82% 82% 82% 82% 82% OR Quartile 2 2.92 2.12 2.85 3.01 2.12 2.90 3.06 2.12 3.00 p Value 3.8E−4 0.0058 0.0012 2.5E−4 0.0058 9.6E−4 2.0E−4 0.0058 6.5E−4 Lower limit of 1.62 1.24 1.51 1.67 1.24 1.54 1.70 1.24 1.60 95% CI Upper limit of 5.26 3.61 5.35 5.44 3.61 5.45 5.53 3.61 5.64 95% CI OR Quartile 3 2.66 2.40 2.91 2.64 2.40 2.80 2.70 2.40 2.94 p Value 3.6E−5 1.3E−4 1.9E−5 3.8E−5 1.3E−4 3.2E−5 2.4E-5 1.3E−4 1.3E−5 Lower limit of 1.67 1.53 1.78 1.66 1.53 1.72 1.70 1.53 1.81 95% CI Upper limit of 4.23 3.75 4.74 4.19 3.75 4.55 4.29 3.75 4.78 95% CI OR Quartile 4 2.43 2.23 3.00 2.34 2.23 2.94 2.46 2.23 3.02 p Value 6.3E−4 0.0020 3.4E−5 0.0010 0.0020 4.7E−5 5.3E-4 0.0020 2.9E−5 Lower limit of 1.46 1.34 1.78 1.41 1.34 1.75 1.48 1.34 1.80 95% CI

Example 12. Use of Metalloproteinase Inhibitor 2 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 2 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 12.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11600 23500 11600 25700 11900 28900 Average 17800 50100 18300 54600 20900 53600 Stdev 21800 77800 22100 82600 35200 78100 p (t-test) 5.7E−8 3.8E−9 5.5E−7 Min 200 2940 200 2940 200 3000 Max 183000 428000 183000 428000 396000 428000 n (Patient) 205 112 221 96 236 81 sCr only Median 11800 21800 11800 25200 12000 24300 Average 19900 46400 20500 49500 23200 45800 Stdev 35400 70300 35500 73900 44300 65800 p (t-test) 1.1E−5 3.7E−6 5.9E−4 Min 200 2940 200 2940 200 2940 Max 428000 396000 428000 396000 428000 371000 n (Patient) 208 108 223 93 235 81 UO only Median 12300 39600 12800 38200 12800 39600 Average 22100 79400 22400 81600 22400 83600 Stdev 34300 102000 34300 106000 34300 106000 p (t-test)  1.3E−11  1.4E−11  4.9E−12 Min 200 5410 200 5410 200 5410 Max 396000 428000 396000 428000 396000 428000 n (Patient) 275 40 278 37 279 36 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.67 0.79 0.71 0.69 0.78 0.72 0.67 0.78 SE 0.032 0.033 0.044 0.033 0.034 0.047 0.035 0.036 0.047 p Value 1.5E−9 3.6E−7  1.3E−10  8.6E−11 1.4E−8 3.6E−9  7.4E−10 2.7E−6 1.2E−9 nCohort Non- 205 208 275 221 223 278 236 235 279 persistent nCohort Persistent 112 108 40 96 93 37 81 81 36 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 85% 83% 90% 89% 87% 89% 90% 86% 89% Specificity 30% 29% 27% 31% 30% 27% 30% 29% 27% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 66% 64% 82% 69% 68% 81% 70% 67% 83% Specificity 59% 57% 55% 58% 57% 54% 57% 56% 54% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 46% 44% 65% 49% 47% 65% 51% 46% 67% Specificity 86% 85% 81% 85% 84% 80% 83% 82% 80% OR Quartile 2 2.42 2.07 3.37 3.43 2.90 3.05 3.93 2.59 2.94 p Value 0.0036 0.015 0.025 4.6E−4 0.0019 0.041 6.0E−4 0.0073 0.049 Lower limit of 95% CI 1.34 1.15 1.16 1.72 1.48 1.04 1.80 1.29 1.01 Upper limit of 95% CI 4.40 3.73 9.81 6.85 5.67 8.89 8.58 5.19 8.60 OR Quartile 3 2.75 2.37 5.66 3.03 2.83 5.02 3.12 2.52 5.90 p Value 3.6E−5 4.3E−4 6.4E−5 1.9E−5 6.3E−5 2.2E−4 3.9E−5 6.2E−4 1.3E−4 Lower limit of 95% CI 1.70 1.46 2.42 1.82 1.70 2.13 1.81 1.48 2.38 Upper limit of 95% CI 4.44 3.82 13.2 5.03 4.71 11.8 5.36 4.28 14.6 OR Quartile 4 5.07 4.24 7.78 5.46 4.82 7.49 5.18 3.86 8.15 p Value 4.0E−9 1.2E−7 1.9E−8 1.0E−9 1.4E−8 8.5E−8 6.3E−9 1.5E−6 4.8E−8 Lower limit of 95% CI 2.95 2.48 3.80 3.17 2.80 3.58 2.97 2.23 3.84 Upper limit of 95% CI 8.72 7.24 15.9 9.42 8.31 15.6 9.02 6.70 17.3

TABLE 12.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11600 21200 11600 21900 11600 24800 Average 17500 45900 17600 50200 20300 49200 Stdev 21300 73300 21200 77700 35600 72600 p (t-test) 9.8E−7 4.0E−8 3.1E−6 Min 200 658 200 2170 200 2170 Max 183000 428000 183000 428000 396000 428000 n (Patient) 186 131 204 113 219 98 sCr only Median 11900 19800 11800 21200 11900 21300 Average 20600 42000 20300 45200 23000 42300 Stdev 36700 66600 35800 69700 45200 61600 p (t-test) 2.7E−4 3.7E−5 0.0020 Min 200 658 200 2170 200 2170 Max 428000 396000 428000 396000 428000 371000 n (Patient) 192 124 206 110 218 98 UO only Median 12000 32200 12500 32200 12300 32400 Average 21600 62900 22000 64300 21900 65300 Stdev 34500 88900 34500 91700 34400 92300 p (t-test) 1.6E−8 1.8E−8 1.0E−8 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 256 59 260 55 261 54 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.63 0.71 0.69 0.66 0.70 0.70 0.66 0.70 SE 0.031 0.032 0.040 0.032 0.033 0.042 0.033 0.034 0.042 p Value 6.8E−8 5.2E−5 1.4E−7 1.2E−9 1.1E−6 1.8E−6  7.7E−10 6.4E−6 1.2E−6 nCohort Non- 186 192 256 204 206 260 219 218 261 persistent nCohort Persistent 131 124 59 113 110 55 98 98 54 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 82% 81% 83% 85% 84% 82% 88% 85% 81% Specificity 30% 29% 27% 30% 30% 27% 31% 29% 26% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 63% 60% 71% 65% 63% 69% 67% 63% 70% Specificity 59% 56% 55% 58% 57% 54% 58% 56% 54% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 41% 39% 53% 45% 42% 53% 47% 42% 54% Specificity 86% 84% 81% 86% 84% 81% 84% 83% 81% OR Quartile 2 2.02 1.67 1.81 2.47 2.15 1.63 3.16 2.30 1.58 p Value 0.012 0.064 0.11 0.0030 0.011 0.20 7.5E−4 0.0087 0.22 Lower limit of 95% CI 1.17 0.971 0.868 1.36 1.20 0.777 1.62 1.23 0.755 Upper limit of 95% CI 3.50 2.88 3.77 4.47 3.87 3.40 6.17 4.28 3.31 OR Quartile 3 2.37 1.90 2.98 2.66 2.21 2.61 2.79 2.19 2.79 p Value 2.3E−4 0.0059 5.0E−4 6.0E−5 0.0010 0.0025 5.6E−5 0.0017 0.0015 Lower limit of 95% CI 1.50 1.20 1.61 1.65 1.38 1.40 1.69 1.34 1.48 Upper limit of 95% CI 3.75 3.01 5.51 4.28 3.56 4.86 4.61 3.57 5.25 OR Quartile 4 4.32 3.28 4.80 4.96 3.77 4.68 4.81 3.41 4.90 p Value 1.2E−7 1.0E−5 3.0E−7 6.1E−9 9.8E−7 7.8E−7 1.1E−8 6.4E−6 4.6E−7 Lower limit of 95% CI 2.51 1.94 2.63 2.89 2.22 2.54 2.81 2.00 2.64 Upper limit of 95% CI 7.41 5.56 8.74 8.52 6.41 8.64 8.26 5.80 9.08

TABLE 12.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11500 21100 11400 21400 11600 21600 Average 16700 44800 16500 48300 19900 46400 Stdev 18500 71600 18100 74800 35100 69800 p (t-test) 9.8E−7 4.3E−8 9.5E−6 Min 200 658 200 2170 200 2170 Max 183000 428000 183000 428000 396000 428000 n (Patient) 175 142 190 127 205 112 sCr only Median 11800 19600 11600 20600 11800 20900 Average 19700 41600 19900 43600 23100 40400 Stdev 35700 65400 36300 67100 46100 59200 p (t-test) 1.6E−4 6.2E−5 0.0044 Min 200 658 200 2170 200 2170 Max 428000 396000 428000 396000 428000 371000 n (Patient) 182 134 195 121 208 108 UO only Median 12000 25100 12000 27000 12000 27000 Average 21200 58500 21500 60100 21500 60100 Stdev 33800 84800 33700 87500 33700 87500 p (t-test) 6.6E−8 6.0E−8 6.0E−8 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 246 69 251 64 251 64 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.64 0.69 0.70 0.66 0.68 0.69 0.65 0.68 SE 0.031 0.032 0.038 0.031 0.032 0.040 0.032 0.033 0.040 p Value 2.5E−8 2.0E−5 4.0E−7  2.2E−10 4.0E−7 4.7E−6 2.6E−9 6.4E−6 4.7E−6 nCohort Non- 175 182 246 190 195 251 205 208 251 persistent nCohort Persistent 142 134 69 127 121 64 112 108 64 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 82% 81% 83% 85% 83% 81% 87% 84% 81% Specificity 31% 29% 27% 32% 30% 27% 31% 30% 27% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 63% 60% 70% 65% 63% 67% 66% 63% 67% Specificity 60% 57% 55% 60% 58% 54% 59% 57% 54% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 39% 38% 48% 43% 40% 48% 43% 40% 48% Specificity 86% 85% 81% 86% 85% 81% 84% 83% 81% OR Quartile 2 2.09 1.71 1.78 2.62 2.19 1.58 2.94 2.27 1.58 p Value 0.0073 0.050 0.099 0.0010 0.0069 0.19 6.5E−4 0.0070 0.19 Lower limit of 1.22 1.00 0.898 1.48 1.24 0.794 1.58 1.25 0.794 95% CI Upper limit of 3.58 2.91 3.52 4.67 3.87 3.14 5.45 4.13 3.14 95% CI OR Quartile 3 2.52 1.98 2.83 2.83 2.33 2.42 2.75 2.23 2.42 p Value 6.9E−5 0.0032 3.6E−4 1.3E−5 3.8E−4 0.0027 3.6E−5 1.0E−3 0.0027 Lower limit of 1.60 1.26 1.60 1.77 1.46 1.36 1.70 1.38 1.36 95% CI Upper limit of 3.97 3.11 5.00 4.51 3.71 4.32 4.44 3.59 4.32 95% CI OR Quartile 4 4.10 3.38 3.99 4.67 3.74 3.97 4.05 3.16 3.97 p Value 4.3E−7 7.4E−6 2.1E−6 2.7E−8 1.2E−6 3.5E−6 2.4E−7 1.9E−5 3.5E−6 Lower limit of 2.37 1.98 2.25 2.71 2.20 2.22 2.38 1.87 2.22 95% CI Upper limit of 7.08 5.76 7.05 8.03 6.37 7.11 6.90 5.35 7.11 95% CI

TABLE 12.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11400 21100 11400 21300 11500 21500 Average 16700 44400 16600 48000 18100 48500 Stdev 18600 71100 18200 74600 23400 76100 p (t-test) 1.3E−6 5.7E−8 2.8E−7 Min 200 658 200 2170 200 2170 Max 183000 428000 183000 428000 215000 428000 n (Patient) 173 144 189 128 201 116 sCr only Median 11700 19400 11500 20400 11600 20900 Average 19700 41300 19200 44100 20600 44300 Stdev 35900 65000 34900 67300 37100 68300 p (t-test) 2.1E−4 2.1E−5 8.1E−5 Min 200 658 200 2170 200 2170 Max 428000 396000 428000 396000 428000 396000 n (Patient) 180 136 192 124 204 112 UO only Median 11900 24000 12000 27000 12000 27000 Average 20900 56100 21300 57800 21300 57800 Stdev 34000 81300 33900 84200 33900 84200 p (t-test) 1.5E-7 1.1E-7 1.1E-7 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 239 76 245 70 245 70 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO UO sCr only UO only UO sCr only UO only UO sCr only only AUC 0.67 0.64 0.70 0.69 0.67 0.69 0.69 0.66 0.69 SE 0.031 0.032 0.037 0.031 0.032 0.038 0.032 0.033 0.038 p Value 1.9E−8 1.9E−5 3.5E−8  2.7E−10 1.3E−7 7.3E−7 2.7E−9 8.0E−7 7.3E−7 nCohort 173 180 239 189 192 245 201 204 245 Non-persistent nCohort 144 136 76 128 124 70 116 112 70 Persistent Cutoff 7990 7980 8020 7990 7980 8020 7990 7980 8020 Quartile 2 Sensitivity 83% 81% 84% 85% 84% 83% 86% 85% 83% Specificity 31% 29% 28% 32% 31% 27% 31% 30% 27% Cutoff 13500 13500 13500 13500 13500 13500 13500 13500 13500 Quartile 3 Sensitivity 62% 60% 70% 65% 63% 67% 66% 63% 67% Specificity 60% 57% 56% 60% 58% 55% 59% 57% 55% Cutoff 28900 28100 29400 28900 28100 29400 28900 28100 29400 Quartile 4 Sensitivity 39% 38% 47% 42% 40% 49% 42% 40% 49% Specificity 86% 84% 82% 86% 85% 82% 85% 83% 82% OR Quartile 2 2.16 1.77 2.08 2.67 2.31 1.82 2.85 2.44 1.82 p Value 0.0050 0.037 0.035 8.3E−4 0.0039 0.086 7.0E−4 0.0034 0.086 Lower limit 1.26 1.03 1.05 1.50 1.31 0.920 1.56 1.34 0.920 of 95% CI Upper limit 3.70 3.01 4.09 4.74 4.07 3.60 5.23 4.43 3.60 of 95% CI OR Quartile 3 2.51 1.97 2.94 2.74 2.37 2.47 2.70 2.33 2.47 p Value 7.1E−5 0.0033 1.3E−4 2.1E−5 2.6E−4 0.0015 4.1E−5 4.7E−4 0.0015 Lower limit 1.59 1.25 1.69 1.72 1.49 1.41 1.68 1.45 1.41 of 95% CI Upper limit 3.96 3.10 5.11 4.37 3.77 4.31 4.34 3.74 4.31 of 95% CI OR Quartile 4 3.95 3.26 4.10 4.57 3.80 4.20 4.01 3.36 4.20 p Value 8.1E−7 1.4E-5 7.2E-7 4.0E-8 9.5E-7 7.9E-7 3.0E-7 6.8E-6 7.9E-7 Lower limit 2.29 1.91 2.35 2.66 2.23 2.38 2.36 1.98 2.38 of 95% CI Upper limit 6.82 5.54 7.17 7.87 6.48 7.42 6.82 5.69 7.42 of 95% CI

TABLE 12.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 168 hours after the sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11400 20900 11300 21400 11400 21600 Average 16600 43600 16500 46900 18100 47600 Stdev 18800 70300 18300 7340 23600 75000 p (t-test) 2.5E-6 1.2E-7 5.2E-7 Min 200 658 200 2170 200 2170 Max 183000 428000 183000 428000 215000 428000 n (Patient) 169 148 184 133 197 120 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11600 19200 11500 20000 11600 20400 Average 19800 40600 19300 43300 20700 43300 Stdev 36300 64200 35200 66400 37400 67300 p (t-test) 3.4E−4 3.9E−5 1.5E−4 Min 200 658 200 2170 200 2170 Max 428000 396000 428000 396000 428000 396000 n (Patient) 176 140 188 128 200 116 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11700 25100 11900 24000 11900 24000 Average 20700 54400 21100 54500 21100 54500 Stdev 34300 79100 34400 80400 34400 80400 p (t-test) 2.6E−7 4.8E−7 4.8E−7 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 234 81 237 78 237 78 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.63 0.71 0.70 0.66 0.70 0.69 0.66 0.70 SE 0.030 0.032 0.035 0.030 0.032 0.036 0.031 0.033 0.036 p Value 2.7E−8 2.8E−5 1.9E−9 1.3E−10 2.1E−7 2.6E−8 1.8E−9 1.4E−6 2.6E−8 nCohort Non- 169 176 234 184 188 237 197 200 237 persistent nCohort Persistent 148 140 81 133 128 78 120 116 78 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 82% 81% 85% 85% 84% 85% 86% 84% 85% Specificity 31% 30% 29% 32% 31% 28% 31% 30% 28% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 62% 59% 72% 65% 62% 71% 66% 63% 71% Specificity 60% 57% 57% 61% 59% 57% 59% 57% 57% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 39% 37% 47% 41% 40% 46% 42% 40% 46% Specificity 86% 85% 82% 86% 85% 82% 85% 84% 82% OR Quartile 2 2.14 1.76 2.31 2.67 2.27 2.17 2.78 2.39 2.17 p Value 0.0051 0.038 0.015 7.0E−4 0.0041 0.025 7.4E−4 0.0036 0.025 Lower limit of 95% CI 1.26 1.03 1.17 1.51 1.30 1.10 1.54 1.33 1.10 Upper limit of 95% CI 3.66 2.98 4.53 4.70 3.98 4.27 5.04 4.29 4.27 OR Quartile 3 2.50 1.96 3.38 2.94 2.35 3.11 2.82 2.30 3.11 p Value 7.3E−5 0.0034 1.3E−5 5.2E−6 2.8E−4 5.3E−5 1.7E−5 5.2E−4 5.3E−5 Lower limit of 95% CI 1.59 1.25 1.95 1.85 1.48 1.79 1.76 1.44 1.79 Upper limit of 95% CI 3.93 3.08 5.85 4.68 3.73 5.39 4.52 3.67 5.39 OR Quartile 4 3.98 3.26 4.16 4.48 3.78 3.87 3.98 3.33 3.87 p Value 8.9E−7 1.5E−5 4.0E−7 6.7E−8 1.1E−6 1.7E−6 3.6E−7 7.9E−6 1.7E−6 Lower limit of 95% CI 2.29 1.91 2.40 2.60 2.22 2.22 2.34 1.96 2.22 Upper limit of 95% CI 6.89 5.56 7.22 7.73 6.46 6.73 6.77 5.63 6.73

TABLE 12.6 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 24 hours after the sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 104000 123000 105000 120000 106000 120000 Average 115000 133000 118000 130000 118000 132000 Stdev 41600 45900 43500 44400 42600 46500 p (t-test) 4.0E−4 0.027 0.014 Min 57000 64800 57000 64800 57000 64800 Max 299000 281000 299000 281000 299000 281000 n (Patient) 206 121 222 105 238 89 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 104000 123000 105000 120000 106000 119000 Average 115000 134000 119000 128000 119000 129000 Stdev 41400 46300 44800 42000 44300 43000 p (t-test) 2.3E−4 0.074 0.082 Min 57000 64800 57000 64800 57000 64800 Max 299000 281000 299000 270000 299000 270000 n (Patient) 208 118 223 103 236 90 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 106000 139000 106000 145000 106000 145000 Average 118000 149000 118000 151000 118000 153000 Stdev 41500 50100 41400 50200 41300 50900 p (t-test) 7.7E−6 2.4E−6 2.0E−6 Min 57000 82200 57000 82200 57000 82200 Max 299000 281000 299000 281000 299000 281000 n (Patient) 281 44 283 42 285 40 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.64 0.71 0.60 0.59 0.72 0.60 0.59 0.72 SE 0.032 0.033 0.046 0.034 0.034 0.047 0.036 0.036 0.048 p Value 2.6E−5 1.6E−5 6.7E−6 0.0036 0.0067 2.1E−6 0.0049 0.017 2.9E−6 nCohort Non- 206 208 281 222 223 283 238 236 285 persistent nCohort Persistent 121 118 44 105 103 42 89 90 40 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 84% 84% 91% 83% 83% 90% 82% 81% 90% Specificity 31% 30% 27% 29% 29% 27% 28% 28% 27% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 63% 64% 73% 60% 59% 76% 61% 59% 78% Specificity 57% 58% 53% 55% 54% 54% 54% 53% 54% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 34% 35% 50% 30% 30% 52% 34% 32% 52% Specificity 80% 80% 79% 77% 77% 79% 78% 78% 79% OR Quartile 2 2.37 2.26 3.77 1.96 1.90 3.55 1.75 1.63 3.33 p Value 0.0032 0.0052 0.014 0.024 0.032 0.019 0.072 0.11 0.027 Lower limit of 95% CI 1.33 1.28 1.31 1.09 1.06 1.23 0.950 0.896 1.15 Upper limit of 95% CI 4.19 4.02 10.9 3.51 3.41 10.3 3.23 2.97 9.67 OR Quartile 3 2.26 2.38 3.05 1.80 1.72 3.71 1.80 1.64 3.99 p Value 5.0E−4 2.6E−4 0.0019 0.015 0.024 5.8E−4 0.021 0.048 4.9E−4 Lower limit of 95% CI 1.43 1.49 1.51 1.12 1.07 1.76 1.09 1.00 1.83 Upper limit of 95% CI 3.59 3.79 6.17 2.88 2.77 7.84 2.95 2.68 8.69 OR Quartile 4 2.06 2.17 3.68 1.51 1.45 4.09 1.82 1.64 4.06 p Value 0.0053 0.0029 9.9E−5 0.12 0.16 3.7E−5 0.029 0.071 5.7E−5 Lower limit of 95% CI 1.24 1.30 1.91 0.895 0.860 2.09 1.06 0.959 2.05 Upper limit of 95% CI 3.43 3.61 7.10 2.54 2.45 7.98 3.11 2.81 8.03

TABLE 12.7 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 48 hours after the sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 103000 122000 104000 121000 104000 123000 Average 115000 131000 117000 130000 116000 133000 Stdev 41800 45200 43100 44400 41900 46200 p (t-test) 5.6E−4 0.0070 0.0013 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 187 140 204 123 219 108 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 103000 123000 104000 120000 105000 120000 Average 115000 132000 117000 130000 117000 131000 Stdev 41500 45800 43100 44800 42100 46400 p (t-test) 4.2E−4 0.013 0.0051 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 192 134 205 121 217 109 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 106000 132000 105000 133000 105000 133000 Average 117000 144000 116000 146000 116000 147000 Stdev 40900 49600 40700 49700 40600 50200 p (t-test) 5.7E−6 1.2E−6 1.1E−6 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 260 65 263 62 265 60 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.63 0.68 0.61 0.60 0.70 0.63 0.61 0.70 SE 0.031 0.032 0.039 0.033 0.033 0.040 0.034 0.034 0.040 p Value 1.6E−5 2.0E−5 3.8E−6 4.5E−4 0.0015 7.0E−7 1.8E−4 0.0014 1.1E−6 nCohort Non- 187 192 260 204 205 263 219 217 265 persistent nCohort Persistent 140 134 65 123 121 62 108 109 60 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 84% 83% 89% 83% 82% 89% 83% 82% 88% Specificity 32% 31% 28% 30% 29% 28% 29% 29% 28% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 61% 62% 68% 60% 59% 71% 61% 59% 72% Specificity 58% 58% 54% 56% 55% 55% 55% 54% 55% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 34% 34% 45% 33% 31% 47% 36% 34% 47% Specificity 81% 81% 80% 79% 79% 80% 80% 79% 80% OR Quartile 2 2.34 2.14 3.30 2.07 1.86 3.08 2.06 1.78 2.93 p Value 0.0021 0.0061 0.0048 0.010 0.027 0.0081 0.015 0.046 0.011 Lower limit of 95% CI 1.36 1.24 1.44 1.19 1.07 1.34 1.15 1.01 1.28 Upper limit of 95% CI 4.04 3.69 7.55 3.62 3.23 7.06 3.70 3.14 6.75 OR Quartile 3 2.23 2.28 2.48 1.91 1.74 2.96 1.94 1.70 3.06 p Value 4.6E−4 3.5E−4 0.0019 0.0052 0.016 3.9E−4 0.0057 0.026 3.4E−4 Lower limit of 95% CI 1.42 1.45 1.40 1.21 1.11 1.62 1.21 1.06 1.66 Upper limit of 95% CI 3.48 3.58 4.41 3.01 2.75 5.39 3.10 2.70 5.63 OR Quartile 4 2.19 2.12 3.15 1.86 1.68 3.48 2.31 1.96 3.42 p Value 0.0024 0.0037 9.3E−5 0.017 0.047 2.7E−5 0.0014 0.010 4.3E−5 Lower limit of 95% CI 1.32 1.28 1.77 1.12 1.01 1.94 1.38 1.17 1.90 Upper limit of 95% CI 3.65 3.52 5.59 3.09 2.79 6.24 3.87 3.29 6.16

Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 72 hours after the sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 100000 120000 103000 120000 103000 122000 Average 114000 131000 116000 130000 115000 133000 Stdev 41600 44900 42800 44300 41600 45900 p (t-test) 2.4E−4 0.0026 3.9E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 176 151 190 137 205 122 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 104000 133000 104000 133000 104000 133000 Average 115000 145000 115000 146000 115000 146000 Stdev 40400 48000 40200 48400 40200 48700 p (t-test) 1.6E−7 4.8E−8 7.4E−8 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 249 76 253 72 254 71 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 104000 133000 104000 133000 104000 133000 Average 115000 145000 115000 146000 115000 146000 Stdev 40400 48000 40200 48400 40200 48700 p (t-test) 1.6E−7 4.8E−8 7.4E−8 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 249 76 253 72 254 71 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.64 0.71 0.62 0.61 0.72 0.64 0.62 0.71 SE 0.031 0.031 0.036 0.032 0.032 0.037 0.032 0.033 0.037 p Value 2.3E−6 3.7E−6 1.1E−8 7.6E−5 3.9E−4 3.7E−9 2.4E−5 3.2E−4 9.2E−9 nCohort Non- 176 182 249 190 194 253 205 207 254 persistent nCohort Persistent 151 144 76 137 132 72 122 119 71 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 83% 83% 91% 82% 82% 90% 83% 82% 90% Specificity 32% 31% 30% 31% 30% 29% 30% 29% 29% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 62% 62% 71% 61% 60% 74% 62% 60% 73% Specificity 60% 60% 56% 58% 57% 57% 57% 56% 56% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 34% 34% 46% 33% 32% 47% 36% 34% 46% Specificity 82% 82% 81% 79% 81% 81% 81% 80% 81% OR Quartile 2 2.41 2.17 4.17 2.07 1.92 3.84 2.04 1.80 3.76 p Value 0.0012 0.0044 6.8E−4 0.0081 0.018 0.0014 0.012 0.037 0.0017 Lower limit of 95% CI 1.42 1.27 1.83 1.21 1.12 1.68 1.17 1.04 1.65 Upper limit of 95% CI 4.11 3.70 9.50 3.54 3.29 8.76 3.56 3.12 8.58 OR Quartile 3 2.50 2.49 3.15 2.18 1.95 3.63 2.20 1.85 3.53 p Value 5.9E−5 6.9E−5 5.1E−5 6.6E−4 0.0035 1.3E−5 7.8E−4 0.0085 2.1E−5 Lower limit of 95% CI 1.60 1.59 1.81 1.39 1.25 2.03 1.39 1.17 1.97 Upper limit of 95% CI 3.90 3.90 5.49 3.41 3.06 6.48 3.48 2.92 6.30 OR Quartile 4 2.39 2.33 3.67 2.02 1.80 3.82 2.48 2.13 3.63 p Value 9.1E−4 0.0012 3.9E−6 0.0064 0.023 2.6E−6 4.9E−4 0.0037 6.5E−6 Lower limit of 95% CI 1.43 1.40 2.11 1.22 1.08 2.18 1.49 1.28 2.07 Upper limit of 95% CI 3.99 3.88 6.37 3.35 2.98 6.69 4.13 3.54 6.37

TABLE 12.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where the persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 99600 120000 103000 120000 103000 123000 Average 113000 132000 115000 131000 114000 134000 Stdev 40000 46100 41400 45700 40300 47100 p (t-test) 4.6E−5 5.8E−4 1.0E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 173 154 188 139 200 127 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 101000 123000 103000 120000 104000 123000 Average 112000 133000 115000 132000 114000 134000 Stdev 39400 46800 41200 46200 40100 47600 p (t-test) 1.8E−5 5.9E−4 1.0E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 179 147 190 136 202 124 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 103000 133000 103000 134000 104000 133000 Average 114000 146000 114000 148000 114000 148000 Stdev 39200 48500 38900 49000 38900 49300 p (t-test) 3.9E−9 7.3E−10 1.2E−9 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 240 85 245 80 246 79 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.66 0.72 0.63 0.63 0.73 0.64 0.64 0.73 SE 0.030 0.031 0.034 0.031 0.032 0.035 0.032 0.032 0.035 p Value 4.6E−7 3.6E−7 5.7E−11 2.1E−5 3.7E−5 1.3E−11 9.2E−6 1.7E−5 3.9E−11 nCohort Non- 173 179 240 188 190 245 200 202 246 persistent nCohort Persistent 154 147 85 139 136 80 127 124 79 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 84% 83% 92% 83% 82% 91% 83% 82% 91% Specificity 33% 32% 31% 31% 31% 30% 30% 30% 30% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 63% 63% 73% 62% 61% 75% 62% 61% 75% Specificity 61% 61% 58% 59% 58% 58% 57% 57% 58% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 34% 35% 47% 33% 33% 49% 36% 36% 48% Specificity 83% 83% 82% 81% 81% 82% 82% 82% 82% OR Quartile 2 2.54 2.28 4.97 2.14 2.05 4.51 2.05 1.96 4.43 p Value 6.2E−4 0.0024 1.3E−4 0.0056 0.0089 3.3E−4 0.011 0.017 3.9E−4 Lower limit of 95% CI 1.49 1.34 2.19 1.25 1.20 1.98 1.18 1.13 1.94 Upper limit of 95% CI 4.32 3.89 11.3 3.66 3.51 10.3 3.55 3.40 10.1 OR Quartile 3 2.69 2.68 3.71 2.29 2.15 4.14 2.23 2.09 4.03 p Value 1.5E−5 1.7E−5 2.2E−6 3.0E−4 8.2E−4 8.9E−7 5.7E−4 0.0015 1.4E−6 Lower limit of 95% CI 1.72 1.71 2.16 1.46 1.37 2.35 1.41 1.33 2.29 Upper limit of 95% CI 4.21 4.21 6.38 3.58 3.37 7.28 3.51 3.30 7.10 OR Quartile 4 2.43 2.54 4.19 2.09 2.04 4.47 2.59 2.54 4.25 p Value 7.5E−4 4.0E−4 2.1E−7 0.0044 0.0056 8.7E−8 2.7E−4 3.5E−4 2.3E−7 Lower limit of 95% CI 1.45 1.52 2.44 1.26 1.23 2.58 1.55 1.52 2.46 Upper limit of 95% CI 4.07 4.24 7.20 3.47 3.39 7.73 4.31 4.23 7.37

TABLE 12.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where the persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 99600 120000 102000 120000 103000 121000 Average 112000 132000 114000 131000 114000 133000 Stdev 39700 46200 41300 45600 40100 47100 p (t-test) 5.0E−5 5.8E−4 1.0E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 169 158 183 144 196 131 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 101000 123000 103000 120000 104000 122000 Average 113000 132000 115000 131000 115000 133000 Stdev 39700 46600 41500 45900 40400 47300 p (t-test) 4.6E−5 0.0013 2.6E−4 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 175 151 186 140 198 128 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 103000 132000 102000 132000 103000 132000 Average 114000 144000 114000 145000 114000 145000 Stdev 39500 48000 39400 48000 39400 48300 p (t-test) 2.0E−8 6.3E−9 1.0E−8 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 235 90 237 88 238 87 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.65 0.72 0.63 0.62 0.72 0.64 0.63 0.72 SE 0.030 0.031 0.034 0.031 0.032 0.034 0.032 0.032 0.034 p Value 8.4E−7 1.2E−6 1.8E−10 2.1E−5 1.0E−4 3.5E−11 1.1E−5 4.9E−5 9.8E−11 nCohort Non- 169 175 235 183 186 237 196 198 238 persistent nCohort Persistent 158 151 90 144 140 88 131 128 87 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 83% 82% 91% 83% 81% 91% 82% 81% 91% Specificity 33% 31% 31% 31% 30% 31% 30% 29% 31% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 63% 63% 72% 62% 61% 74% 62% 61% 74% Specificity 62% 61% 58% 59% 58% 59% 58% 57% 58% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 34% 34% 44% 33% 32% 45% 36% 35% 45% Specificity 83% 82% 82% 81% 80% 82% 82% 81% 82% OR Quartile 2 2.34 2.10 4.62 2.15 1.89 4.45 2.02 1.80 4.37 p Value 0.0015 0.0054 1.1E−4 0.0048 0.018 1.7E−4 0.011 0.033 2.0E−4 Lower limit of 95% CI 1.39 1.25 2.12 1.26 1.11 2.05 1.17 1.05 2.01 Upper limit of 95% CI 3.96 3.56 10.0 3.67 3.20 9.68 3.48 3.08 9.51 OR Quartile 3 2.68 2.67 3.63 2.33 2.14 4.01 2.21 2.07 3.91 p Value 1.5E−5 1.8E−5 1.8E−6 2.1E−4 8.5E−4 5.0E−7 6.1E−4 0.0016 8.2E−7 Lower limit of 95% CI 1.72 1.70 2.14 1.49 1.37 2.33 1.40 1.32 2.27 Upper limit of 95% CI 4.20 4.18 6.17 3.64 3.35 6.89 3.47 3.26 6.72 OR Quartile 4 2.44 2.37 3.68 2.05 1.91 3.87 2.57 2.36 3.68 p Value 7.6E−4 0.0010 1.7E−6 0.0056 0.012 7.5E−7 2.9E−4 9.6E−4 1.9E−6 Lower limit of 95% CI 1.45 1.42 2.16 1.23 1.15 2.26 1.54 1.42 2.16 Upper limit of 95% CI 4.09 3.96 6.27 3.40 3.16 6.61 4.29 3.93 6.30

Example 13. Use of Metalloproteinase Inhibitor 2 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 2 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 13.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where the persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11300 18000 11500 19500 11400 20900 Average 15600 37400 16300 42100 17400 44700 Stdev 13600 63200 14600 69300 20800 71900 p (t-test) 2.5E−4 6.9E−6 2.1E−6 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 119 198 158 159 179 138 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 12000 16000 11900 19200 11900 19600 Average 17300 37100 16800 41900 18500 43600 Stdev 14800 64900 14600 70200 22500 72900 p (t-test) 7.0E−4 1.2E−5 1.6E−5 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 130 186 162 154 184 132 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11600 31200 12000 32600 12000 35500 Average 20100 59500 20800 66400 21300 70300 Stdev 33400 81800 33200 89300 33300 93700 p (t-test) 4.6E−9 3.6E−10 1.4E−10 Min 200 5410 200 5410 200 5410 Max 396000 428000 396000 428000 396000 428000 n (Patient) 241 74 256 59 263 52 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.59 0.76 0.65 0.63 0.76 0.67 0.64 0.76 SE 0.031 0.032 0.035 0.031 0.031 0.038 0.031 0.032 0.041 p Value 2.1E−5 0.0066 6.1E−14 1.5E−6 5.4E−5 8.5E−12 5.2E−8 7.3E−6 9.5E−11 nCohort Non- 119 130 241 158 162 256 179 184 263 persistent nCohort Persistent 198 186 74 159 154 59 138 132 52 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 80% 78% 93% 82% 81% 92% 85% 84% 90% Specificity 34% 30% 31% 32% 30% 29% 32% 32% 28% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 56% 53% 76% 60% 58% 76% 63% 61% 77% Specificity 60% 55% 58% 59% 57% 56% 60% 58% 55% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 33% 32% 51% 36% 36% 54% 39% 38% 58% Specificity 87% 85% 83% 85% 85% 82% 85% 84% 81% OR Quartile 2 2.06 1.56 6.11 2.08 1.79 4.39 2.67 2.43 3.68 p Value 0.0060 0.087 1.8E−4 0.0063 0.028 0.0024 5.9E−4 0.0019 0.0078 Lower limit of 95% CI 1.23 0.937 2.37 1.23 1.06 1.69 1.53 1.39 1.41 Upper limit of 95% CI 3.46 2.61 15.8 3.50 3.02 11.4 4.68 4.26 9.62 OR Quartile 3 1.89 1.37 4.24 2.18 1.85 4.07 2.54 2.09 4.10 p Value 0.0070 0.17 1.6E−6 6.6E−4 0.0071 2.2E−5 6.6E−5 0.0015 6.1E−5 Lower limit of 95% CI 1.19 0.873 2.35 1.39 1.18 2.13 1.61 1.33 2.06 Upper limit of 95% CI 2.99 2.15 7.64 3.41 2.88 7.78 4.00 3.30 8.16 OR Quartile 4 3.39 2.56 5.15 3.28 3.19 5.27 3.78 3.26 5.96 p Value 1.1E−4 0.0012 1.4E−8 2.2E−5 2.9E−5 6.3E−8 1.3E−6 1.2E−5 3.1E−8 Lower limit of 95% CI 1.83 1.45 2.92 1.90 1.85 2.89 2.21 1.92 3.17 Upper limit of 95% CI 6.28 4.51 9.07 5.68 5.51 9.62 6.48 5.54 11.2

TABLE 13.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where the persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 10600 17900 11400 19300 11300 20000 Average 15200 36900 16000 40700 17100 42500 Stdev 13700 62200 14300 67300 21100 69000 p (t-test) 3.0E−4 1.7E−5 9.1E−6 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 112 205 147 170 165 152 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11800 15700 11800 18800 11600 19300 Average 17200 36800 16800 40600 17800 41800 Stdev 14900 64200 14700 68700 21200 70000 p (t-test) 8.5E-4 3.4E-5 2.9E-5 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 126 190 154 162 169 147 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11400 22900 11700 22900 11900 27000 Average 19900 52100 20700 56200 21200 58100 Stdev 34300 74900 34000 80800 34100 83900 p (t-test) 2.9E-7 9.4E-8 8.4E-8 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 222 93 238 77 245 70 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.59 0.73 0.65 0.62 0.72 0.67 0.64 0.71 SE 0.031 0.032 0.033 0.031 0.031 0.036 0.030 0.031 0.038 p Value 1.6E−6 0.0039 1.9E−12 7.9E−7 1.1E−4 1.3E−9 1.7E−8 9.4E−6 1.8E−8 nCohort Non- 112 126 222 147 154 238 165 169 245 persistent nCohort Persistent 205 190 93 170 162 77 152 147 70 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 81% 79% 89% 81% 80% 87% 84% 82% 86% Specificity 36% 31% 31% 32% 30% 29% 33% 31% 28% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 57% 54% 7% 60% 57% 70% 62% 59% 70% Specificity 62% 56% 59% 61% 58% 56% 61% 58% 56% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 33% 32% 46% 35% 35% 47% 38% 37% 49% Specificity 88% 85% 84% 86% 85% 82% 86% 85% 82% OR Quartile 2 2.36 1.68 3.74 2.03 1.66 2.74 2.67 2.13 2.35 p Value 0.0012 0.048 3.0E−4 0.0075 0.052 0.0062 4.0E−4 0.0056 0.021 Lower limit of 95% CI 1.40 1.01 1.83 1.21 0.995 1.33 1.55 1.25 1.14 Upper limit of 95% CI 3.98 2.81 7.65 3.40 2.79 5.63 4.59 3.63 4.86 OR Quartile 3 2.22 1.45 3.45 2.37 1.85 3.03 2.63 2.00 2.91 p Value 9.5E−4 0.11 3.2E−6 1.8E−4 0.0071 8.3E−5 3.0E−5 0.0024 2.4E−4 Lower limit of 95% CI 1.38 0.921 2.05 1.51 1.18 1.74 1.67 1.28 1.65 Upper limit of 95% CI 3.55 2.28 5.82 3.72 2.88 5.25 4.14 3.14 5.15 OR Quartile 4 3.70 2.60 4.44 3.19 3.01 3.98 3.70 3.34 4.20 p Value 7.6E−5 0.0012 6.8E−8 4.9E−5 8.3E−5 1.1E−6 3.0E−6 1.2E−5 7.9E−7 Lower limit of 95% CI 1.93 1.46 2.58 1.82 1.74 2.28 2.14 1.95 2.38 Upper limit of 95% CI 7.07 4.62 7.64 5.58 5.21 6.95 6.42 5.75 7.42

TABLE 13.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 10600 18000 11400 19300 11400 19300 Average 15200 36900 16000 40300 17300 41100 Stdev 13700 62000 14400 66800 21400 67800 p (t-test) 3.3E−4 2.3E−5 3.4E−5 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 111 206 144 173 158 159 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11500 16000 11600 18600 11600 18800 Average 17200 36600 16900 40100 18100 40600 Stdev 15000 63900 14800 68100 21500 68900 p (t-test) 9.8E−4 5.3E−5 8.5E−5 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 124 192 151 165 163 153 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11400 21700 11600 21700 11800 21700 Average 20000 48700 20700 51100 21200 52200 Stdev 34800 72100 34700 76000 34700 78500 p (t-test) 2.8E−6 1.8E−6 2.0E−6 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 212 103 225 90 232 83 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.60 0.71 0.65 0.62 0.70 0.65 0.63 0.69 SE 0.031 0.032 0.033 0.031 0.031 0.034 0.031 0.031 0.036 p Value 1.2E−6 0.0030 1.8E−10 8.5E−7 1.7E−4 1.3E−8 5.0E−7 6.1E−5 1.7E−7 nCohort Non- 111 124 212 144 151 225 158 163 232 persistent nCohort Persistent 206 192 103 173 165 90 159 153 83 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 81% 79% 87% 81% 79% 86% 82% 81% 84% Specificity 36% 31% 31% 32% 30% 29% 32% 31% 28% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 57% 54% 68% 60% 58% 68% 61% 59% 67% Specificity 63% 56% 58% 62% 58% 57% 61% 58% 56% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 33% 31% 43% 35% 34% 43% 36% 35% 45% Specificity 88% 85% 83% 86% 85% 82% 86% 85% 82% OR Quartile 2 2.41 1.74 3.13 1.99 1.64 2.46 2.23 1.89 2.14 p Value 9.3E−4 0.034 5.8E−4 0.0089 0.060 0.0070 0.0029 0.017 0.023 Lower limit of 1.43 1.04 1.63 1.19 0.979 1.28 1.32 1.12 1.11 95% CI Upper limit of 4.06 2.92 6.00 3.34 2.73 4.73 3.78 3.19 4.13 95% CI OR Quartile 3 2.29 1.53 2.99 2.44 1.90 2.78 2.42 2.00 2.64 p Value 6.2E−4 0.066 1.5E−5 1.2E−4 0.0051 1.0E−4 1.2E−4 0.0025 3.0E−4 Lower limit of 1.42 0.972 1.82 1.55 1.21 1.66 1.54 1.28 1.56 95% CI Upper limit of 3.68 2.41 4.91 3.84 2.96 4.64 3.80 3.12 4.48 95% CI OR Quartile 4 3.63 2.51 3.77 3.29 2.86 3.54 3.55 3.01 3.64 p Value 9.5E−5 0.0017 1.0E−6 3.8E−5 1.7E−4 4.4E−6 7.4E−6 6.3E−5 3.7E−6 Lower limit of 1.90 1.41 2.21 1.87 1.65 2.06 2.04 1.75 2.11 95% CI Upper limit of 6.95 4.47 6.43 5.80 4.95 6.06 6.18 5.17 6.29 95% CI

TABLE 13.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 10600 17900 11300 19300 11300 19300 Average 15300 36700 16000 40200 17300 40800 Stdev 13700 61900 14400 66700 21500 67500 p (t-test) 4.0E−4 2.7E−5 4.3E−5 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 110 207 143 174 156 161 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11500 16000 11600 18600 11500 18700 Average 17200 36600 16900 39900 18100 40400 Stdev 15000 63900 14900 68000 21600 68700 p (t-test) 9.8E−4 6.2E−5 1.0E−4 Min 200 658 200 658 200 658 Max 76000 428000 76000 428000 215000 428000 n (Patient) 124 192 150 166 162 154 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11400 21700 11600 21500 11700 21500 Average 20200 47300 20800 49200 21300 49800 Stdev 35100 71100 35000 74400 35200 76400 p (t-test) 8.5E−6 5.9E−6 8.6E−6 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 208 107 220 95 226 89 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.60 0.69 0.65 0.62 0.69 0.65 0.63 0.68 SE 0.031 0.032 0.033 0.031 0.031 0.034 0.031 0.031 0.035 p Value 3.2E−6 0.0030 3.4E−9 9.4E−7 1.8E−4 4.8E−8 3.8E−7 6.6E−5 4.3E−7 nCohort Non- 110 124 208 143 150 220 156 162 226 persistent nCohort Persistent 207 192 107 174 166 95 161 154 89 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 81% 79% 86% 81% 80% 85% 83% 81% 84% Specificity 35% 31% 31% 32% 30% 30% 33% 31% 29% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 57% 54% 66% 60% 57% 66% 61% 58% 66% Specificity 63% 56% 58% 62% 58% 57% 61% 58% 56% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 32% 31% 41% 34% 34% 42% 36% 35% 43% Specificity 88% 85% 83% 86% 85% 82% 86% 85% 82% OR Quartile 2 2.29 1.74 2.73 2.03 1.66 2.43 2.31 1.92 2.16 p Value 0.0018 0.034 0.0015 0.0074 0.052 0.0064 0.0019 0.014 0.018 Lower limit of 1.36 1.04 1.47 1.21 0.995 1.28 1.36 1.14 1.14 95% CI Upper limit of 3.86 2.92 5.07 3.40 2.78 4.59 3.91 3.25 4.10 95% CI OR Quartile 3 2.23 1.53 2.74 2.38 1.85 2.59 2.42 1.94 2.52 p Value 9.1E−4 0.066 4.8E−5 1.8E−4 0.0071 2.0E−4 1.2E−4 0.0036 4.0E−4 Lower limit of 1.39 0.972 1.69 1.51 1.18 1.57 1.54 1.24 1.51 95% CI Upper limit of 3.59 2.41 4.46 3.74 2.89 4.28 3.80 3.04 4.21 95% CI OR Quartile 4 3.57 2.51 3.45 3.24 2.81 3.38 3.43 2.96 3.36 p Value 1.2E−4 0.0017 4.5E−6 4.8E−5 2.2E−4 8.1E−6 1.3E−5 8.1E−5 1.1E−5 Lower limit of 1.87 1.41 2.03 1.84 1.62 1.98 1.97 1.73 1.96 95% CI Upper limit of 6.83 4.47 5.86 5.70 4.86 5.76 5.97 5.08 5.77 95% CI

TABLE 13.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE critetia. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 10700 17900 11300 19300 11300 19300 Average 14900 36400 15500 39800 17000 40300 Stdev 12600 61500 13600 65900 21300 66500 p (t-test) 4.3E−4 2.6E−5 5.1E−5 Min 200 658 200 658 200 658 Max 61200 428000 61200 428000 215000 428000 n (Patient) 106 211 138 179 150 167 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11600 16000 11600 18600 11500 18700 Average 16800 36500 16500 39700 17700 40200 Stdev 14200 63600 14100 67300 21300 68000 p (t-test) 8.9E−4 5.4E−5 9.1E−5 Min 200 658 200 658 200 658 Max 63200 428000 63200 428000 215000 428000 n (Patient) 121 195 146 170 158 158 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 11200 21700 11400 21700 11500 21700 Average 19700 46600 20400 47800 21000 47900 Stdev 35400 69400 35400 71800 35700 73200 p (t-test) 7.4E−6 8.7E−6 1.5E−5 Min 200 2170 200 2170 200 2170 Max 396000 428000 396000 428000 396000 428000 n (Patient) 202 113 212 103 217 98 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.60 0.71 0.65 0.62 0.70 0.66 0.63 0.69 SE 0.032 0.032 0.032 0.030 0.031 0.033 0.030 0.031 0.034 p Value 8.4E−6 0.0031 3.8E−11 4.1E−7 1.1E−4 1.3E−9 3.0E−7 4.2E−5 1.1E−8 nCohort Non- 106 121 202 138 146 212 150 158 217 persistent nCohort Persistent 211 195 113 179 170 103 167 158 98 Cutoff Quartile 2 7990 7980 8020 7990 7980 8020 7990 7980 8020 Sensitivity 80% 79% 87% 81% 79% 86% 83% 81% 86% Specificity 35% 31% 32% 33% 30% 31% 33% 31% 30% Cutoff Quartile 3 13500 13500 13500 13500 13500 13500 13500 13500 13500 Sensitivity 56% 54% 68% 60% 57% 68% 60% 58% 68% Specificity 62% 56% 60% 62% 58% 58% 61% 58% 58% Cutoff Quartile 4 28900 28100 29400 28900 28100 29400 28900 28100 29400 Sensitivity 32% 31% 42% 35% 34% 42% 36% 35% 42% Specificity 89% 85% 84% 87% 86% 83% 87% 85% 82% OR Quartile 2 2.16 1.72 3.03 2.06 1.66 2.81 2.38 1.92 2.57 p Value 0.0040 0.039 4.5E−4 0.0059 0.052 0.0014 0.0012 0.014 0.0037 Lower limit of 1.28 1.03 1.63 1.23 0.996 1.49 1.41 1.14 1.36 95% CI Upper limit of 3.64 2.88 5.63 3.46 2.78 5.30 4.02 3.23 4.85 95% CI OR Quartile 3 2.13 1.50 3.20 2.46 1.85 2.99 2.43 1.94 2.99 p Value 0.0019 0.083 2.7E−6 1.1E−4 0.0070 1.5E−5 1.2E−4 0.0036 2.0E−5 Lower limit of 1.32 0.948 1.97 1.56 1.18 1.82 1.54 1.24 1.81 95% CI Upper limit of 3.44 2.36 5.19 3.88 2.90 4.91 3.81 3.04 4.95 95% CI OR Quartile 4 3.72 2.60 3.78 3.53 3.08 3.50 3.64 3.22 3.39 p Value 1.1E−4 0.0013 9.3E−7 2.2E−5 8.3E−5 3.7E−6 7.8E−6 3.0E−5 7.1E−6 Lower limit of 1.91 1.45 2.22 1.97 1.76 2.06 2.07 1.86 1.99 95% CI Upper limit of 7.25 4.68 6.44 6.33 5.40 5.96 6.43 5.58 5.77 95% CI

TABLE 13.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (OU) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 98000 117000 98700 119000 102000 120000 Average 109000 129000 113000 129000 116000 129000 Stdev 37500 45800 42900 43700 44400 42800 p (t-test) 8.6E−5 9.6E−4 0.012 Min 57600 57000 57000 64800 57000 64800 Max 277000 299000 299000 281000 299000 281000 n (Patient) 114 213 155 172 179 148 sCr only Median 98000 120000 98700 123000 103000 122000 Average 107000 131000 112000 131000 117000 128000 Stdev 35400 46600 41700 44300 45900 41000 p (t-test) 2.4E−6 6.5E−5 0.025 Min 57600 57000 57000 64800 57000 64800 Max 277000 299000 299000 281000 299000 270000 n (Patient) 124 202 157 169 182 144 UO only Median 106000 125000 106000 129000 105000 132000 Average 118000 134000 118000 140000 117000 145000 Stdev 42500 46600 41900 48100 41600 48700 p (t-test) 0.0039 2.4E−4 1.6E−5 Min 57000 70200 57000 70200 57000 77400 Max 299000 281000 299000 281000 299000 281000 n (Patient) 245 80 261 64 269 56 Persistence Period Duration (hr) 24 48 72 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.65 0.67 0.62 0.64 0.66 0.66 0.62 0.62 0.69 SE 0.031 0.030 0.037 0.030 0.030 0.040 0.031 0.031 0.042 p Value 4.4E−7 4.0E−9 0.0013 2.8E−6 6.2E−8 9.2E−5 1.6E−4 2.2E−4 5.8E−6 nCohort Non- 114 124 245 155 157 261 179 182 269 persistent nCohort Persistent 213 202 80 172 169 64 148 144 56 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 80% 80% 85% 83% 83% 89% 83% 82% 91% Specificity 34% 34% 28% 34% 34% 28% 32% 31% 28% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 58% 60% 62% 61% 63% 66% 63% 63% 71% Specificity 65% 66% 54% 62% 64% 54% 60% 60% 54% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 31% 33% 38% 31% 33% 41% 31% 32% 45% Specificity 87% 88% 79% 81% 83% 79% 80% 80% 79% OR Quartile 2 2.06 2.07 2.22 2.56 2.46 3.22 2.30 2.02 4.02 p Value 0.0058 0.0049 0.020 3.8E−4 6.7E−4 0.0057 0.0022 0.0093 0.0044 Lower limit 1.23 1.25 1.13 1.52 1.46 1.41 1.35 1.19 1.54 of 95% CI Upper limit 3.43 3.45 4.36 4.31 4.13 7.39 3.92 3.42 10.4 of 95% CI OR Quartile 3 2.58 2.92 1.95 2.55 2.95 2.21 2.57 2.62 2.97 p Value 8.2E−5 6.8E−6 0.012 3.9E−5 2.5E−6 0.0065 3.6E−5 2.7E−5 6.8E−4 Lower limit 1.61 1.83 1.16 1.63 1.88 1.25 1.64 1.67 1.58 of 95% CI Upper limit 4.13 4.65 3.27 3.98 4.63 3.91 4.03 4.12 5.56 of 95% CI OR Quartile 4 3.03 3.61 2.23 1.94 2.50 2.50 1.79 1.90 3.00 p Value 4.2E−4 4.2E−5 0.0041 0.012 7.0E−4 0.0019 0.024 0.013 3.5E−4 Lower limit 1.64 1.95 1.29 1.15 1.47 1.40 1.08 1.15 1.64 of 95% CI Upper limit 5.60 6.66 3.85 3.25 4.24 4.47 2.97 3.16 5.48 of 95% CI

TABLE 13.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (OU) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 97100 118000 98500 120000 99700 120000 Average 106000 130000 112000 129000 115000 129000 Stdev 33300 46500 42700 43700 44300 42800 p (t-test) 2.8E−6 5.3E−4 0.0050 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 108 219 144 183 164 163 sCr only Median 97500 120000 98700 121000 100000 121000 Average 105000 132000 111000 131000 115000 129000 Stdev 31200 47500 41100 44700 43800 43300 p (t-test) 8.4E−8 6.2E−5 0.0032 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 120 206 149 177 166 160 UO only Median 104000 124000 104000 128000 104000 130000 Average 116000 135000 116000 139000 116000 142000 Stdev 42300 45600 41600 46700 41300 47200 p (t-test) 5.8E−4 3.7E−5 5.6E−6 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 225 100 241 84 248 77 Persistence Period Duration (hr) 24 48 72 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.68 0.69 0.64 0.65 0.66 0.67 0.63 0.63 0.69 SE 0.030 0.029 0.034 0.030 0.030 0.036 0.031 0.031 0.037 p Value 2.2E−9 6.2E−11 4.3E−5 5.8E−7 9.0E−8 2.3E−6 2.2E−5 2.1E−5 4.1E−7 nCohort Non- 108 120 225 144 149 241 164 166 248 persistent nCohort Persistent 219 206 100 183 177 84 163 160 77 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 80% 81% 86% 83% 82% 89% 83% 82% 90% Specificity 36% 35% 30% 35% 34% 30% 33% 32% 29% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 59% 61% 64% 61% 62% 67% 63% 62% 70% Specificity 69% 68% 56% 64% 64% 56% 62% 62% 56% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 32% 33% 38% 31% 33% 40% 31% 32% 43% Specificity 89% 89% 80% 83% 84% 80% 81% 81% 80% OR Quartile 2 2.31 2.23 2.60 2.51 2.29 3.55 2.37 2.12 3.60 p Value 0.0014 0.0020 0.0030 4.4E−4 0.0015 8.5E−4 0.0012 0.0045 0.0013 Lower limit 1.38 1.34 1.38 1.50 1.37 1.69 1.41 1.26 1.65 of 95% CI Upper limit 3.87 3.72 4.91 4.19 3.82 7.47 3.99 3.56 7.86 of 95% CI OR Quartile 3 3.18 3.33 2.26 2.79 2.97 2.50 2.75 2.72 2.99 p Value 3.3E−6 7.1E−7 9.9E−4 8.4E−6 2.4E−6 5.4E−4 9.3E−6 1.2E−5 9.0E−5 Lower limit 1.95 2.07 1.39 1.78 1.89 1.49 1.76 1.74 1.73 of 95% CI Upper limit 5.18 5.36 3.68 4.38 4.68 4.21 4.30 4.26 5.18 of 95% CI OR Quartile 4 3.76 4.15 2.52 2.15 2.54 2.73 1.95 2.04 3.05 p Value 9.3E−5 1.5E−5 5.1E−4 0.0048 6.9E−4 2.5E−4 0.010 0.0065 7.0E−5 Lower limit 1.93 2.18 1.50 1.26 1.48 1.60 1.17 1.22 1.76 of 95% CI Upper limit 7.30 7.90 4.25 3.67 4.35 4.68 3.26 3.40 5.27 of 95% CI

TABLE 13.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (OU) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 97000 118000 98300 119000 99100 120000 Average 106000 130000 112000 129000 114000 129000 Stdev 33500 46400 43000 43500 44100 42900 p (t-test) 2.8E−6 6.3E−4 0.0036 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 107 220 142 185 158 169 sCr only Median 97100 120000 98500 121000 99100 121000 Average 104000 132000 111000 131000 113000 130000 Stdev 31100 47300 41300 44400 42900 43700 p (t-test) 4.6E−8 5.1E−5 5.6E−4 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 118 208 146 180 160 166 UO only Median 104000 123000 104000 125000 104000 127000 Average 115000 135000 115000 138000 115000 141000 Stdev 41900 45600 41400 46400 41000 46900 p (t-test) 1.6E−4 1.6E−5 2.7E−6 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 216 109 230 95 237 88 Persistence Period Duration (hr) 24 48 72 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.68 0.70 0.65 0.65 0.66 0.67 0.63 0.65 0.69 SE 0.030 0.029 0.033 0.030 0.030 0.034 0.030 0.030 0.035 p Value 1.4E−9 8.6E−12 6.1E−6 4.8E−7 2.8E−8 4.7E−7 1.1E−5 1.4E−6 8.8E−8 nCohort Non- 107 118 216 142 146 230 158 160 237 persistent nCohort Persistent 220 208 109 185 180 95 169 166 88 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 80% 81% 86% 83% 82% 89% 83% 83% 90% Specificity 36% 36% 31% 35% 34% 31% 34% 33% 30% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 60% 61% 64% 61% 62% 66% 62% 63% 69% Specificity 69% 69% 57% 64% 65% 57% 63% 63% 57% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 32% 34% 38% 31% 33% 39% 31% 33% 41% Specificity 89% 90% 81% 82% 84% 80% 81% 82% 81% OR Quartile 2 2.36 2.32 2.76 2.60 2.41 3.80 2.44 2.34 3.83 p Value 0.0011 0.0012 0.0013 2.7E−4 7.8E−4 2.4E−4 7.6E−4 0.0013 4.0E−4 Lower limit 1.41 1.39 1.49 1.55 1.44 1.86 1.45 1.39 1.82 of 95% CI Upper limit 3.95 3.87 5.11 4.34 4.02 7.74 4.09 3.93 8.05 of 95% CI OR Quartile 3 3.30 3.57 2.37 2.80 3.07 2.56 2.75 2.87 2.99 p Value 1.8E−6 2.1E−7 3.6E−4 8.2E−6 1.3E−6 2.2E−4 9.3E−6 4.2E−6 3.8E−5 Lower limit 2.02 2.21 1.48 1.78 1.95 1.55 1.76 1.83 1.78 of 95% CI Upper limit 5.39 5.78 3.82 4.40 4.83 4.22 4.31 4.50 5.03 of 95% CI OR Quartile 4 3.69 4.48 2.57 2.08 2.61 2.62 1.90 2.27 2.87 p Value 1.1E−4 9.2E−6 3.3E−4 0.0069 5.4E−4 3.2E−4 0.015 0.0020 1.0E−4 Lower limit 1.90 2.31 1.54 1.22 1.51 1.55 1.13 1.35 1.69 of 95% CI Upper limit 7.18 8.69 4.31 3.55 4.49 4.44 3.17 3.83 4.90 of 95% CI

TABLE 13.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (OU) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 96500 118000 98000 120000 98700 120000 Average 106000 129000 111000 130000 113000 129000 Stdev 33600 46300 41000 44600 42500 44100 p (t-test) 3.6E−6 1.1E−4 9.9E−4 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 106 221 140 187 155 172 sCr only Median 97100 120000 98300 122000 98800 122000 Average 104000 132000 110000 131000 112000 131000 Stdev 31100 47300 39100 45500 41100 44900 p (t-test) 4.6E−8 7.0E−6 1.0E−4 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 118 208 144 182 158 168 UO only Median 104000 124000 103000 127000 103000 129000 Average 114000 136000 114000 139000 114000 141000 Stdev 40300 47300 39900 47600 39700 48000 p (t-test) 2.3E−5 8.8E−7 1.7E−7 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 211 114 223 102 229 96 Persistence Period Duration (hr) 24 48 72 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.68 0.70 0.66 0.66 0.67 0.69 0.64 0.65 0.70 SE 0.030 0.029 0.033 0.030 0.029 0.033 0.030 0.030 0.034 p Value 2.0E−9 8.6E−12 1.9E−6 7.3E−8 3.3E−9 2.1E−8 2.9E−6 2.9E−7 4.8E−9 nCohort Non- 106 118 211 140 144 223 155 158 229 persistent nCohort Persistent 221 208 114 187 182 102 172 168 96 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 81% 81% 86% 83% 82% 89% 83% 83% 90% Specificity 37% 36% 31% 36% 35% 31% 34% 34% 31% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 59% 61% 64% 61% 63% 68% 62% 63% 70% Specificity 69% 69% 57% 65% 66% 58% 63% 64% 58% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 32% 34% 38% 31% 33% 40% 31% 33% 42% Specificity 89% 90% 82% 83% 85% 82% 81% 83% 82% OR Quartile 2 2.41 2.32 2.73 2.69 2.49 3.78 2.56 2.42 3.86 p Value 8.5E−4 0.0012 0.0011 1.6E−4 4.8E−4 1.4E−4 3.8E−4 8.4E−4 2.0E−4 Lower limit 1.44 1.39 1.49 1.61 1.49 1.90 1.52 1.44 1.90 of 95% CI Upper limit 4.04 3.87 4.99 4.50 4.17 7.52 4.31 4.06 7.88 of 95% CI OR Quartile 3 3.22 3.57 2.39 2.97 3.25 2.87 2.83 3.03 3.20 p Value 3.0E−6 2.1E−7 2.7E−4 2.9E−6 4.3E−7 2.7E−5 5.6E−6 1.5E−6 7.4E−6 Lower limit 1.97 2.21 1.50 1.88 2.06 1.75 1.81 1.93 1.92 of 95% CI Upper limit 5.26 5.78 3.83 4.68 5.13 4.70 4.43 4.76 5.32 of 95% CI OR Quartile 4 3.63 4.48 2.67 2.17 2.73 2.98 1.94 2.36 3.18 p Value 1.4E−4 9.2E−6 1.8E−4 0.0047 3.4E−4 3.9E−5 0.012 0.0013 1.6E−5 Lower limit 1.87 2.31 1.60 1.27 1.57 1.77 1.15 1.40 1.88 of 95% CI Upper limit 7.06 8.69 4.47 3.72 4.72 5.02 3.25 3.99 5.38 of 95% CI

TABLE 13.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 96500 118000 97900 119000 98600 120000 Average 105000 129000 111000 130000 113000 129000 Stdev 32700 46400 40700 44700 42500 44000 p (t-test) 3.3E−96 1.0E−94 8.3E−94 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 102 225 135 192 149 178 sCr only Median 97200 120000 98500 120000 99100 120000 Average 105000 131000 110000 130000 113000 130000 Stdev 31400 47200 39500 45500 41400 44900 p (t-test) 1.6E−97 3.3E−95 4.1E−94 Min 57600 57000 57600 57000 57600 57000 Max 252000 299000 299000 281000 299000 281000 n (Patient) 115 211 140 186 154 172 UO only Median 104000 123000 103000 125000 103000 125000 Average 115000 134000 114000 137000 114000 138000 Stdev 40800 46800 40500 46800 40200 47400 p (t-test) 1.3E−94 5.9E−96 3.3E−96 Min 57600 57000 57600 57000 57600 57000 Max 299000 281000 299000 281000 299000 281000 n (Patient) 204 121 214 111 219 106 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.68 0.69 0.64 0.66 0.66 0.67 0.64 0.64 0.68 SE 0.030 0.029 0.032 0.030 0.030 0.032 0.030 0.030 0.033 p Value 3.3E−99 1.0E−910 1.2E−95 7.8E−98 7.2E−98 8.5E−98 1.9E−96 3.4E−96 1.0E−97 nCohort Non-persistent 102 115 204 135 140 214 149 154 219 nCohort Persistent 225 211 121 192 186 111 178 172 106 Cutoff Quartile 2 90400 90300 90600 90400 90300 90600 90400 90300 90600 Sensitivity 80% 80% 84% 82% 81% 87% 83% 81% 87% Specificity 36% 35% 30% 36% 34% 31% 34% 32% 31% Cutoff Quartile 3 109000 109000 109000 109000 109000 109000 109000 109000 109000 Sensitivity 59% 60% 63% 61% 61% 67% 62% 62% 68% Specificity 69% 69% 57% 65% 65% 58% 64% 63% 58% Cutoff Quartile 4 137000 137000 137000 137000 137000 137000 137000 137000 137000 Sensitivity 32% 33% 36% 31% 32% 38% 31% 32% 39% Specificity 89% 90% 81% 84% 84% 81% 82% 82% 81% OR Quartile 2 2.28 2.15 2.34 2.56 2.18 3.16 2.47 2.10 2.90 p Value 0.0019 0.0034 0.0036 3.1E−94 0.0026 3.5E−94 5.8E−94 0.0044 9.6E−94 Lower limit of 95% CI 1.35 1.29 1.32 1.54 1.31 1.68 1.48 1.26 1.54 Upper limit of 95% CI 3.83 3.58 4.16 4.28 3.62 5.93 4.13 3.51 5.45 OR Quartile 3 3.10 3.32 2.27 2.92 2.94 2.81 2.85 2.73 2.98 p Value 7.4E−96 1.0E−96 4.9E−94 4.4E−96 3.5E−96 2.4E−95 5.4E−96 1.1E−95 1.2E−95 Lower limit of 95% CI 1.89 2.05 1.43 1.85 1.86 1.74 1.81 1.74 1.83 Upper limit of 95% CI 5.10 5.37 3.60 4.62 4.64 4.54 4.47 4.28 4.85 OR Quartile 4 3.81 4.26 2.33 2.33 2.55 2.65 2.02 2.21 2.74 p Value 1.3E−94 1.8E−95 0.0011 0.0025 8.2E−94 2.1E−94 0.0086 0.0030 1.4E−94 Lower limit of 95% CI 1.92 2.20 1.40 1.35 1.47 1.58 1.20 1.31 1.63 Upper limit of 95% CI 7.57 8.27 3.89 4.04 4.42 4.43 3.41 3.74 4.60

Example 14. Use of Metalloproteinase Inhibitor 4 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 4 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 14.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11.1 29.5 11.8 29.0 11.1 28.9 Average 17.9 190 18.2 183 17.1 180 Stdev 21.6 850 21.6 834 22.3 826 p (t-test) 0.13 0.18 0.21 Min 0.00110 0.00167 0.00167 0.00110 0.00167 0.00110 Max 108 9030 108 9030 108 9030 n (Patient) 57 260 46 271 41 276 sCr only Median 13.0 29.0 13.3 28.8 13.3 28.8 Average 22.6 203 23.1 201 21.3 199 Stdev 27.4 884 27.6 880 21.5 873 p (t-test) 0.077 0.083 0.091 Min 0.00110 0.00167 0.00167 0.00110 0.00167 0.00110 Max 173 9030 173 9030 108 9030 n (Patient) 76 240 74 242 70 246 UO only Median 17.0 38.8 16.9 34.6 15.7 33.7 Average 97.5 261 98.2 236 98.8 220 Stdev 655 931 691 863 725 819 p (t-test) 0.069 0.12 0.16 Min 0.00110 0.00277 0.00110 0.00273 0.00110 0.00273 Max 9030 7570 9030 7570 9030 7570 n (Patient) 194 121 173 142 156 159 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.71 0.66 0.66 0.69 0.65 0.66 0.70 0.65 0.66 SE 0.034 0.034 0.032 0.038 0.034 0.031 0.038 0.035 0.030 p Value 1.3E−99 3.4E−96 4.9E−97 9.4E−97 1.9E−95 5.3E−97 1.2E−97 1.8E−95 7.3E−98 nCohort Recovered 57 76 194 46 74 173 41 70 156 nCohort Non-recovered 260 240 121 271 242 142 276 246 159 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 80% 78% 88% 78% 77% 86% 78% 77% 85% Specificity 46% 33% 33% 43% 32% 34% 46% 31% 35% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 55% 55% 64% 54% 54% 62% 55% 54% 62% Specificity 74% 64% 59% 74% 64% 60% 80% 64% 62% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 30% 31% 36% 29% 31% 33% 28% 31% 33% Specificity 96% 95% 81% 96% 95% 82% 95% 96% 83% OR Quartile 2 3.28 1.69 3.48 2.76 1.63 3.16 3.11 1.52 3.06 p Value 1.1E−94 0.070 7.7E−95 0.0022 0.093 7.2E−95 0.0010 0.16 5.6E−95 Lower limit of 95% CI 1.79 0.958 1.88 1.44 0.921 1.79 1.58 0.847 1.78 Upper limit of 95% CI 5.98 2.97 6.45 5.30 2.89 5.57 6.12 2.73 5.28 OR Quartile 3 3.48 2.18 2.58 3.36 2.05 2.40 4.98 2.12 2.71 p Value 1.3E−94 0.0042 7.3E−95 6.9E−94 0.0085 1.6E−94 9.8E−95 0.0074 1.8E−95 Lower limit of 95% CI 1.84 1.28 1.62 1.67 1.20 1.52 2.22 1.22 1.72 Upper limit of 95% CI 6.58 3.72 4.13 6.77 3.51 3.78 11.2 3.67 4.28 OR Quartile 4 11.8 8.18 2.42 8.89 7.86 2.18 7.68 9.98 2.32 p Value 7.6E−94 7.8E−95 8.5E−94 0.0030 1.1E−94 0.0033 0.0057 1.5E−94 0.0019 Lower limit of 95% CI 2.80 2.88 1.44 2.10 2.77 1.30 1.81 3.04 1.37 Upper limit of 95% CI 49.5 23.2 4.07 37.6 22.3 3.66 32.6 32.7 3.95

TABLE 14.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 11.2 33.4 12.3 29.7 11.2 29.7 Average 21.2 214 22.6 205 21.7 199 Stdev 26.7 908 27.7 887 28.6 873 p (t-test) 0.045 0.070 0.089 Min 0.00110 0.00167 0.00167 0.00110 0.00167 0.00110 Max 121 9030 121 9030 121 9030 n (Patient) 90 227 79 238 71 246 sCr only Median 12.5 33.7 12.7 33.4 12.7 33.6 Average 25.3 227 25.7 225 24.4 221 Stdev 32.9 940 33.1 936 30.2 926 p (t-test) 0.028 0.031 0.036 Min 0.00110 0.00167 0.00167 0.00110 0.00167 0.00110 Max 173 9030 173 9030 172 9030 n (Patient) 106 211 104 213 99 218 UO only Median 18.2 33.4 18.5 30.7 17.2 33.2 Average 111 262 117 229 109 226 Stdev 645 986 674 913 690 871 p (t-test) 0.10 0.21 0.18 Min 0.00110 0.00277 0.00110 0.00273 0.00110 0.00273 Max 9030 7570 9030 7570 9030 7570 n (Patient) 212 103 194 121 178 137 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.71 0.68 0.64 0.68 0.67 0.61 0.69 0.67 0.62 SE 0.030 0.030 0.034 0.032 0.031 0.033 0.032 0.031 0.032 p Value 2.8E−12 7.0E−99 7.9E−95 2.9E−98 5.4E−98 0.0012 2.0E−99 3.6E−98 1.9E−94 nCohort Recovered 90 106 212 79 104 194 71 99 178 nCohort Non-recovered 227 211 103 238 213 121 246 218 137 Cutoff Quartile 2 8.33 8.33 8.36 8.33 8.33 8.36 8.33 8.33 8.36 Sensitivity 83% 81% 89% 81% 81% 86% 81% 80% 85% Specificity 44% 37% 32% 42% 37% 32% 45% 36% 33% Cutoff Quartile 3 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 Sensitivity 58% 57% 62% 56% 57% 59% 57% 57% 60% Specificity 70% 64% 56% 68% 63% 55% 73% 65% 57% Cutoff Quartile 4 60.9 60.9 61.8 60.9 60.9 61.8 60.9 60.9 61.8 Sensitivity 33% 34% 33% 31% 33% 30% 30% 33% 31% Specificity 93% 92% 79% 92% 91% 78% 93% 92% 79% OR Quartile 2 3.86 2.49 3.95 2.99 2.42 2.87 3.47 2.33 2.90 p Value 9.7E−97 6.5E−94 9.2E−95 9.5E−95 9.9E−94 5.1E−94 1.6E−95 0.0017 2.4E−94 Lower limit of 95% CI 2.25 1.47 1.98 1.73 1.43 1.58 1.97 1.37 1.64 Upper limit of 95% CI 6.62 4.20 7.86 5.19 4.08 5.21 6.11 3.94 5.12 OR Quartile 3 3.24 2.41 2.06 2.78 2.28 1.75 3.61 2.41 2.00 p Value 1.0E−95 3.5E−94 0.0033 2.0E−94 7.9E−94 0.017 1.6E−95 4.5E−94 0.0027 Lower limit of 95% CI 1.92 1.49 1.27 1.62 1.41 1.10 2.02 1.48 1.27 Upper limit of 95% CI 5.47 3.89 3.33 4.77 3.70 2.76 6.47 3.94 3.15 OR Quartile 4 6.77 5.47 1.83 5.49 5.28 1.49 5.79 5.61 1.68 p Value 1.8E−95 6.9E−96 0.025 1.4E−94 1.1E−95 0.13 2.9E−94 1.3E−95 0.046 Lower limit of 95% CI 2.83 2.61 1.08 2.29 2.52 0.887 2.24 2.58 1.01 Upper limit of 95% CI 16.2 11.5 3.10 13.2 11.1 2.49 15.0 12.2 2.81

TABLE 14.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.3 33.7 12.3 33.5 12.3 33.4 Average 24.0 235 24.5 228 24.1 219 Stdev 32.3 958 32.9 942 33.8 922 p (t-test) 0.019 0.026 0.038 Min 0.00110 0.00167 0.00167 0.00110 0.00167 0.00110 Max 223 9030 223 9030 223 9030 n (Patient) 114 203 107 210 97 220 sCr only Median 12.7 35.5 13.3 33.4 13.3 34.0 Average 27.3 248 27.8 244 26.9 239 Stdev 36.2 989 36.5 982 34.6 970 p (t-test) 0.013 0.015 0.018 Min 0.00110 0.00167 0.00167 0.00110 0.00167 0.00110 Max 223 9030 223 9030 223 9030 n (Patient) 127 190 124 193 119 198 UO only Median 18.1 33.4 17.7 33.4 16.9 33.7 Average 112 263 115 243 106 243 Stdev 642 996 658 952 669 910 p (t-test) 0.11 0.16 0.12 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 214 101 204 111 190 125 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.70 0.68 0.62 0.69 0.66 0.61 0.69 0.66 0.64 SE 0.029 0.030 0.035 0.030 0.030 0.034 0.030 0.030 0.032 p Value 4.9E−12 4.5E−9 4.7E−4 3.2E−10 1.1E−7 9.4E−4 2.2E−10 9.3E−8 1.1E−5 nCohort Recovered 114 127 214 107 124 204 97 119 190 nCohort Non-recovered 203 190 101 210 193 111 220 198 125 Cutoff Quartile 2 8.33 8.33 8.36 8.33 8.33 8.36 8.33 8.33 8.36 Sensitivity 84% 82% 86% 82% 81% 85% 82% 81% 86% Specificity 40% 35% 30% 39% 35% 30% 40% 34% 32% Cutoff Quartile 3 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 Sensitivity 60% 59% 62% 60% 58% 62% 60% 58% 65% Specificity 68% 63% 56% 68% 62% 56% 71% 63% 59% Cutoff Quartile 4 60.9 60.9 61.8 60.9 60.9 61.8 60.9 60.9 61.8 Sensitivity 34% 35% 32% 33% 35% 30% 33% 34% 32% Specificity 91% 90% 78% 91% 90% 77% 92% 90% 79% OR Quartile 2 3.48 2.82 2.71 3.02 2.32 2.41 3.03 2.21 2.81 p Value 3.6E−6 4.9E−4 0.0021 3.8E−5 0.0015 0.0038 4.4E−5 0.0026 5.3E−4 Lower limit of 95% CI 2.05 1.50 1.44 1.79 1.38 1.33 1.78 1.32 0.57 Upper limit of 95% CI 5.91 4.23 5.12 5.11 3.88 4.38 5.15 3.71 5.04 OR Quartile 3 3.13 2.44 2.08 3.16 2.27 2.12 3.63 2.36 2.70 p Value 3.4E−6 1.5E−4 0.0031 4.5E−6 5.2E−4 0.0018 9.5E−7 3.1E−4 3.1E−5 Lower limit of 95% CI 1.94 1.54 1.28 1.93 1.43 1.32 2.17 1.48 1.69 Upper limit of 95% CI 5.08 3.88 3.37 5.16 3.60 3.41 6.07 3.77 4.31 OR Quartile 4 5.47 4.78 1.65 4.85 4.54 1.45 5.41 4.66 1.82 p Value 2.7E−6 2.1E−6 0.065 1.4E−5 4.5E−6 0.16 2.0E−5 5.6E−6 0.022 Lower limit of 95% CI 2.69 2.50 0.970 2.38 2.38 0.861 2.49 2.40 1.09 Upper limit of 95% CI 11.1 9.12 2.80 9.88 8.67 2.45 11.8 9.07 3.05

TABLE 14.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 12.3   37.0 12.3   36.1 11.9   33.7 Average 26.8   256 27.5   245 27.6   233 Stdev 42.8   1010 44.0   984 45.6   958 p (t-test) 0.0088 0.014 0.023 Min  0.00110 0.00167  0.00167 0.00110  0.00167 0.00110 Max 337     9030 337    9030 337    9030 n (Patient) 134     183 125    192 114    203 sCr only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 12.5   37.0 12.7   36.7 12.7   36.6 Average 28.8   263 29.1   260 28.2   257 Stdev 43.9   1020 44.1   1020 42.8   1010 p (t-test) 0.0073 0.0082 0.0090 Min  0.00110 0.00167  0.00110 0.00167  0.00110 0.00167 Max 337    9030 337    9030 337    9030 n (Patient) 140    177 138    179 135    182 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 18.1  38.8 17.3  36.3 17.0  35.8 Average 110    270 113    251 104    251 Stdev 640    1000 653    964 662    924 p (t-test) 0.089 0.13 0.10 Min   0.00110 0.00277   0.00110 0.00277   0.00110 0.00277 Max 9030     7570 9030     7570 9030     7570 n (Patient) 216    99 207    108 194    121 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC  0.71  0.69  0.64  0.70  0.68  0.63  0.70  0.69  0.65 SE  0.028  0.029  0.035  0.029  0.030  0.034  0.029  0.029  0.032 p Value 6.0E−14 5.9E−11 7.2E−5 9.9E−12 5.2E−10 8.2E−5 2.5E−11 2.4E−10 5.1E−6 nCohort 134    140    216    125    138    207    114    135    194    Recovered nCohort Non- 183    177    99   192    179    108    203    182    121    recovered Cutoff Quartile 2  8.33  8.33  8.36  8.33  8.33  8.36  8.33  8.33  8.36 Sensitivity 85% 84% 86% 83% 83% 86% 83% 83% 86% Specificity 38% 36% 30% 38% 36% 31% 39% 36% 32% Cutoff Quartile 3 24.1  24.1  24.1  24.1  24.1  24.1  24.1  24.1  24.1  Sensitivity 62% 60% 64% 61% 60% 64% 61% 60% 65% Specificity 66% 63% 56% 67% 62% 57% 69% 63% 59% Cutoff Quartile 4 60.9  60.9  61.8  60.9  60.9  61.8  60.9  60.9  61.8  Sensitivity 37% 37% 34% 35% 36% 32% 34% 36% 33% Specificity 90% 89% 79% 90% 89% 79% 90% 90% 80% OR Quartile 2  3.40  2.84  2.61  3.01  2.73  2.77  3.02  2.69  2.87 p Value 6.6E−6  1.1E−4    0.0031 3.8E−5  1.7E−4  0.0013 3.6E−5  2.1E−4  5.1E−4 Lower limit of  2.00  1.67  1.38  1.78  1.62  1.49  1.79  1.59  1.58 95% CI Upper limit of  5.79  4.80  4.94  5.09  4.62  5.16  5.10  4.53  5.21 95% CI OR Quartile 3  3.27  2.59  2.23  3.27  2.46  2.35  3.54  2.54  2.74 p Value 6.6E−7 4.5E−5   0.0013 9.4E−7 1.1E−4 4.9E−4 3.7E−7 6.7E−5 2.8E−5 Lower limit of  2.05  1.64  1.37  2.04  1.56  1.45  2.18  1.61  1.71 95% CI Upper limit of  5.21  4.08  3.64  5.24  3.88  3.79  5.77  4.01  4.39 95% CI OR Quartile 4  5.38  4.84  1.99  4.62  4.68  1.78  4.82  4.92  1.96 p Value 3.4E−7  5.4E−7   0.011 3.5E−6  9.4E−7   0.031 7.0E−6  7.4E−7    0.011 Lower limit of  2.82  2.61  1.17  2.42  2.52  1.05  2.43  2.62  1.17 95% CI Upper limit of 10.3   8.96  3.37  8.81  8.66  3.00  9.58  9.24  3.29 95% CI

TABLE 14.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 16.8  37.0 16.5  36.9 16.5  36.9 Average 101    244 102    239 102    239 Stdev 672    896 677    886 677    886 p (t-test) 0.11 0.12 0.12 Min   0.00110 0.00169   0.00110 0.00169   0.00110 0.00169 Max 9030     7570 9030     7570 9030     7570 n (Patient) 188    129 185    132 185    132 sCr only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 15.1   38.6 15.4   37.8 15.1   38.6 Average 36.5   284 36.8   280 35.9   279 Stdev 67.1   1080 67.5   1070 66.9   1070 p (t-test) 0.0041 0.0050 0.0050 Min  0.00110 0.00167  0.00110 0.00167  0.00110 0.00167 Max 453    9030 453    9030 453    9030 n (Patient) 160    157 157    160 156    161 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 18.2  39.1 18.5  37.0 18.1  37.9 Average 104    275 105    268 105    262 Stdev 643    985 647    972 652    959 p (t-test) 0.065 0.077 0.086 Min    0.00277   0.00110 0.00277   0.00110 0.00277 Max 9030     7570 9030     7570 9030     7570 n (Patient) 211    104 208    107 205    110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC  0.65  0.68  0.64  0.65  0.67  0.63  0.65  0.68  0.64 SE  0.032  0.030  0.034  0.032  0.030  0.034  0.032  0.030  0.033 p Value 3.1E−6 1.5E−9 3.4E−5 2.5E−6 8.1E−9 6.6E−5 2.5E−6 2.3E−9 2.3E−5 nCohort 188    160    211    185    157    208    185    156    205    Recovered nCohort Non- 129    157    104    132    160    107    132    161    110    recovered Cutoff Quartile 2  8.33  8.33  8.36  8.33  8.33  8.36  8.33  8.33  8.36 Sensitivity 84% 84% 86% 85% 83% 86% 85% 83% 86% Specificity 31% 34% 30% 32% 33% 31% 32% 33% 31% Cutoff Quartile 3 24.1  24.1  24.1  24.1  24.1  24.1  24.1  24.1  24.1  Sensitivity 63% 62% 63% 63% 62% 63% 63% 62% 64% Specificity 59% 62% 56% 59% 62% 56% 59% 62% 57% Cutoff Quartile 4 60.9  60.9  61.8  60.9  60.9  61.8  60.9  60.9  61.8  Sensitivity 37% 38% 36% 37% 38% 35% 37% 38% 35% Specificity 83% 87% 80% 83% 87% 80% 83% 88% 80% OR Quartile 2  2.49  2.69  2.58  2.62  2.44  2.73  2.62  2.48  2.87 p Value   0.0016 3.2E−4   0.0027 8.7E−4 9.9E−4   0.0015 8.7E−4 7.9E−4 8.4E−4 Lower limit of  1.41  1.57  1.39  1.49  1.43  1.47  1.49  1.46  1.55 95% CI Upper limit of  4.40  4.61  4.81  4.62  4.15  5.07  4.62  4.22  5.34 95% CI OR Quartile 3  2.38  2.70  2.25  2.43  2.62  2.15  2.43  2.70  2.33 p Value 2.2E−4 1.9E−5   0.0010 1.5E−4 3.0E−5   0.0017 1.5E−4 1.9E−5 5.2E−4 Lower limit of  1.50  1.71  1.39  1.54  1.67  1.34  1.54  1.71  1.44 95% CI Upper limit of  3.77  4.24  3.64  3.84  4.13  3.47  3.84  4.24  3.75 95% CI OR Quartile 4  2.89  3.98  2.22  2.93  4.11  2.09  2.93  4.40  2.11 p Value 6.7E−5 1.4E−6   0.0029 5.5E−5 1.1E−6   0.0057 5.5E−5 4.6E−7   0.0049 Lower limit of  1.71  2.27  1.31  1.74  2.33  1.24  1.74  2.47  1.25 95% CI Upper limit of  4.87  6.98  3.76  4.95  7.25  3.52  4.95  7.82  3.55 95% CI

TABLE 14.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 3180 3320 3180 3320 3180 3300 Average 3270 3980 3200 3970 3220 3950 Stdev 1390 2560 1240 2540 1270 2530 p (t-test) 0.050 0.057 0.079 Min 1080 863 1080 863 1080 863 Max 7920 19700 5620 19700 5620 19700 n (Patient) 52 275 42 285 38 289 sCr only Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 3140 3320 3130 3330 3130 3320 Average 3340 4000 3240 4020 3280 4000 Stdev 1570 2590 1380 2610 1390 2600 p (t-test) 0.046 0.018 0.035 Min 1080 863 1080 863 1080 863 Max 9820 19700 7920 19700 7920 19700 n (Patient) 69 257 67 259 63 263 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 3010 3910 2980 3860 3030 3770 Average 3430 4530 3380 4430 3390 4320 Stdev 1930 2890 1940 2770 1990 2680 p (t-test) 4.5E−5 8.0E−5 4.8E−4 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 197 128 176 149 159 166 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.56 0.56 0.62 0.56 0.57 0.62 0.56 0.56 0.61 SE 0.042 0.038 0.032 0.046 0.038 0.031 0.048 0.039 0.031 p V alue 0.13 0.12 2.5E−4 0.16 0.062 1.5E−4 0.22 0.12 5.5E−4 nCohort Recovered 52 69 197 42 67 176 38 63 159 nCohort Non- 275 257 128 285 259 149 289 263 166 recovered Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 76% 76% 83% 76% 76% 82% 76% 76% 82% Specificity 33% 30% 30% 33% 31% 31% 32% 30% 32% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 51% 51% 60% 51% 51% 59% 51% 51% 57% Specificity 52% 52% 56% 52% 54% 57% 53% 54% 57% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 27% 28% 32% 27% 28% 31% 26% 27% 30% Specificity 85% 84% 79% 86% 85% 80% 84% 84% 79% OR Quartile 2 1.57 1.41 2.06 1.60 1.48 2.00 1.44 1.37 2.14 p Value 0.17 0.26 0.010 0.19 0.19 0.0098 0.33 0.31 0.0039 Lower limit of 95% CI 0.825 0.781 1.19 0.795 0.821 1.18 0.692 0.746 1.28 Upper limit of 95% CI 2.98 2.53 3.57 3.20 2.67 3.38 3.01 2.52 3.59 OR Quartile 3 1.10 1.12 1.95 1.12 1.21 1.94 1.13 1.22 1.70 p Value 0.74 0.68 0.0038 0.73 0.49 0.0033 0.72 0.48 0.017 Lower limit of 95% CI 0.610 0.656 1.24 0.587 0.704 1.25 0.576 0.702 1.10 Upper limit of 95% CI 2.00 1.90 3.06 2.15 2.07 3.02 2.23 2.11 2.64 OR Quartile 4 2.02 2.01 1.79 2.18 2.19 1.74 1.90 2.00 1.60 p Value 0.084 0.050 0.024 0.090 0.034 0.032 0.17 0.062 0.070 Lower limit of 95% CI 0.911 0.999 1.08 0.884 1.06 1.05 0.766 0.965 0.962 Upper limit of 95% CI 4.50 4.05 2.97 5.38 4.53 2.88 4.73 4.14 2.66

TABLE 14.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 3040 3390 3070 3350 3040 3390 Average 3300 4070 3330 4030 3290 4020 Stdev 1510 2660 1560 2610 1600 2580 p (t-test) 0.011 0.026 0.026 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 9100 19700 n (Patient) 87 240 76 251 69 258 sCr only Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 3040 3390 3040 3440 2980 3430 Average 3380 4090 3310 4110 3330 4090 Stdev 1640 2680 1520 2700 1530 2680 p (t-test) 0.015 0.0059 0.011 Min 1080 863 1080 863 1080 863 Max 9820 19700 9100 19700 9100 19700 n (Patient) 101 226 99 228 94 233 UO only Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 3060 3890 3050 3720 3070 3720 Average 3520 4540 3510 4400 3490 4330 Stdev 2070 2880 2090 2760 2120 2670 p (t-test) 2.9E−4 0.0011 0.0018 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 217 108 198 127 181 144 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.57 0.61 0.57 0.58 0.60 0.58 0.57 0.60 SE 0.035 0.034 0.034 0.036 0.034 0.033 0.037 0.034 0.032 p Value 0.026 0.037 9.7E−4 0.055 0.019 0.0023 0.030 0.030 0.0019 nCohort Recovered 87 101 217 76 99 198 69 94 181 nCohort Non- 240 226 108 251 228 127 258 233 144 recovered Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 78% 78% 82% 78% 78% 80% 78% 78% 81% Specificity 33% 32% 29% 34% 32% 28% 35% 32% 30% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 52% 53% 60% 52% 53% 58% 53% 53% 57% Specificity 56% 55% 55% 57% 57% 55% 59% 57% 55% Cutoff Quartile 4 4720 4720 4720 4720 4720 4720 4720 4720 4720 Sensitivity 29% 29% 33% 28% 29% 32% 28% 28% 31% Specificity 85% 83% 79% 84% 84% 79% 84% 83% 80% OR Quartile 2 1.76 1.63 1.87 1.81 1.70 1.61 1.84 1.63 1.84 p Value 0.039 0.067 0.033 0.037 0.048 0.082 0.038 0.071 0.023 Lower limit of 95% CI 1.03 0.967 1.05 1.03 1.01 0.942 1.03 0.958 1.09 Upper limit of 95% CI 3.03 2.76 3.33 3.17 2.87 2.75 3.27 2.78 3.12 OR Quartile 3 1.43 1.38 1.84 1.42 1.47 1.71 1.63 1.54 1.63 p Value 0.16 0.18 0.011 0.18 0.11 0.020 0.075 0.082 0.029 Lower limit of 95% CI 0.870 0.864 1.15 0.848 0.916 1.09 0.952 0.947 1.05 Upper limit of 95% CI 2.33 2.22 2.93 2.39 2.37 2.68 2.80 2.49 2.54 OR Quartile 4 2.30 1.99 1.86 2.06 2.11 1.83 2.00 1.93 1.77 p Value 0.012 0.023 0.018 0.036 0.016 0.020 0.052 0.035 0.027 Lower limit of 95% CI 1.20 1.10 1.11 1.05 1.15 1.10 0.994 1.05 1.07 Upper limit of 95% CI 4.41 3.62 3.11 4.05 3.88 3.03 4.03 3.54 2.93

TABLE 14.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-rccovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO 24 48 72 Recovery Period Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Duration (hr) Cohort Cohort Cohort Cohort Cohort Cohort Median 3040 3470 3100 3450 2990 3470 Average 3390 4120 3430 4080 3390 4060 Stdev 1820 2650 1870 2620 1940 2580 p (t-test) 0.0097 0.024 0.024 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 111 216 104 223 94 233 sCr only 24 48 72 Recovery Period Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Duration (hr) Cohort Cohort Cohort Cohort Cohort Cohort Median 2990 3490 2950 3500 2930 3490 Average 3380 4170 3340 4180 3350 4160 Stdev 1880 2670 1800 2680 1820 2660 p (t-test) 0.0046 0.0023 0.0037 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 124 203 121 206 116 211 UO only 24 48 72 Recovery Period Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Duration (hr) Cohort Cohort Cohort Cohort Cohort Cohort Median 3090 3710 3100 3700 3040 3770 Average 3540 4530 3550 4420 3480 4420 Stdev 2060 2910 2080 2830 2080 2740 p (t-test) 4.8lf-4 0.0017 5.5 If-4 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 219 106 208 117 193 132 Recovery Period 24 48 72 Duration (hr) sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.58 0.59 0.60 0.57 0.60 0.59 0.58 0.60 0.61 SE 0.033 0.032 0.034 0.033 0.032 0.033 0.034 0.032 0.032 p Value 0.013 0.0032 0.0029 0.034 0.0020 0.0070 0.013 0.0028 8.2E-4 nCohort Recovered 111 124 219 104 121 208 94 116 193 nCohort Non-recovered 216 203 106 223 206 117 233 211 132 Cutoff Quartile 2 2320 2320 2330 2320 2320 2,330 2,320 2,320 2,330 Sensitivity 79% 80% 83% 78% 80% 81% 79% 80% 82% Specificity 32% 34% 29% 33% 34% 28% 34% 34% 30% Cutoff Quartile 3 3280 3280 3280 3280 3280 3,280 3,280 3,280 3,280 Sensitivity 53% 54% 58% 52% 54% 56% 54% 55% 58% Specificity 56% 56% 53% 55% 57% 53% 59% 58% 55% Cutoff Quartile 4 4720 4720 4720 4720 4720 4,720 4,720 4,720 4,720 Sensitivity Specificity 29% 30% 31% 28% 30% 30% 27% 29% 31% 83% 83% 78% 82% 83% 77% 81% 83% 79% OR Quartile 2 1.77 2.09 1.97 1.77 2.06 1.71 1.89 1.98 1.89 p Value 0.029 0.0045 0.023 0.031 0.0053 0.057 0.018 0.0088 0.021 Lower limit of 95% CI 1.06 1.26 1.10 1.05 1.24 0.983 1.11 1.19 1.10 Upper limit of 95% CI 2.97 3.47 3.55 2.98 3.43 2.97 3.21 3.30 3.24 OR Quartile 3 1.44 1.53 1.55 1.34 1.58 1.48 1.63 1.64 1.65 p Value 0.12 0.063 0.064 0.22 0.047 0.091 0.047 0.034 0.027 Lower limit of 95% CI 0.909 0.978 0.974 0.839 1.01 0.939 1.01 1.04 1.06 Upper limit of 95% CI 2.28 2.40 2.48 2.14 2.49 2.34 2.65 2.59 2.58 OR Quartile 4 1.99 2.11 1.57 1.76 2.17 1.46 1.60 2.00 1.67 p Value 0.019 0.0087 0.090 0.054 0.0070 0.15 0.12 0.017 0.046 Lower limit of 95% CI 1.12 1.21 0.933 0.990 1.24 0.876 0.887 1.13 1.01 Upper limit of 95% CI 3.54 3.68 2.64 3.14 3.82 2.44 2.88 3.52 2.77

TABLE 14.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-rccovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO 24 48 72 Recovery Period Duration (hr) Recovered Cohort Non- recovered Cohort Recovered Cohort Non- recovered Cohort Recovered Cohort Non- recovered Cohort Median 3150 3430 3160 3350 3040 3470 Average 3450 4160 3500 4100 3430 4100 Stdev 1790 2750 1830 2710 1910 2640 p (t-test) 0.0098 0.028 0.018 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 134 193 125 202 113 214 sCr only 24 48 72 Recovery Period Duration (hr) Recovered Cohort Non- recovered Cohort Recovered Cohort Non- recovered Cohort Recovered Cohort Non- recovered Cohort Median 3130 3450 3130 3450 3110 3470 Average 3470 4170 3430 4190 3430 4170 Stdev 1860 2740 1790 2760 1800 2740 p (t-test) 0.0099 0.0054 0.0061 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 139 188 137 190 133 194 UO only 24 48 72 Recovery Period Duration (hr) Recovered Cohort Non- recovered Cohort Recovered Cohort Non- recovered Cohort Recovered Cohort Non- recovered Cohort Median 3080 3720 3090 3620 3020 3860 Average 3510 4600 3520 4480 3470 4470 Stdev 2030 2940 2050 2870 2070 2760 p (t-test) 1.31*-4 5.7E−4 2.2E−4 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 220 105 211 114 198 127 Recovery Period 24 48 72 Duration (hr) sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.56 0.57 0.61 0.55 0.57 0.60 0.58 0.58 0.62 SE 0.032 0.032 0.034 0.032 0.032 0.033 0.033 0.032 0.032 p Value 0.044 0.026 0.0012 0.10 0.021 0.0034 0.016 0.017 2.9E−4 nCohort Recovered 134 139 220 125 137 211 113 133 198 nCohort Non-recovered 193 188 105 202 190 114 214 194 127 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 78% 79% 84% 78% 79% 83% 79% 79% 83% Specificity 29% 31% 29% 30% 31% 29% 33% 31% 30% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 52% 52% 57% 51% 52% 56% 54% 53% 58% Specificity 52% 53% 53% 51% 53% 53% 57% 53% 55% Cutoff Quartile 4 4720 4720 4720 4720 4720 4720 4720 4720 4720 Sensitivity 30% 30% 32% 28% 30% 31% 28% 29% 31% Specificity 81% 81% 78% 80% 82% 78% 80% 81% 79% OR Quartile 2 1.43 1.71 2.12 1.47 1.66 2.08 1.83 1.66 2.19 p Value 0.16 0.037 0.013 0.14 0.049 0.012 0.021 0.048 0.0057 Lower limit of 95% CI 0.865 1.03 1.17 0.883 1.00 1.17 1.10 1.00 1.26 Upper limit of 95% CI 2.37 2.83 3.85 2.44 2.74 3.70 3.05 2.75 3.83 OR Quartile 3 1.18 1.20 1.51 1.09 1.21 1.45 1.52 1.27 1.71 p Value 0.47 0.41 0.082 0.70 0.41 0.11 0.075 0.29 0.020 Lower limit of 95% CI 0.757 0.776 0.948 0.699 0.776 0.916 0.959 0.816 1.09 Upper limit of 95% CI 1.83 1.87 2.42 1.71 1.87 2.29 2.40 1.98 2.68 OR Quartile 4 1.83 1.84 1.72 1.57 1.92 1.55 1.49 1.80 1.71 sCr or UO p Value 0.027 0.023 0.041 0.097 0.016 0.096 0.15 0.031 0.038 Lower limit of 95% CI 1.07 1.09 1.02 0.921 1.13 0.925 0.862 1.05 1.03 Upper limit of 95% CI 3.12 3.13 2.88 2.68 3.27 2.58 2.57 3.06 2.83

TABLE 14.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-rccovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO 24 48 72 Recovery Period Duration (hr) Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Median 3090 3510 3080 3520 3070 3620 Average 3520 4330 3520 4320 3510 4320 Stdev 2090 2750 2110 2730 2110 2720 p (t-test) 0.0027 0.0028 0.0027 Min 1080 863 1080 863 1080 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 187 140 184 143 183 144 sCr only 24 48 72 Recovery Period Duration (hr) Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Median 3130 3450 3140 3430 3140 3430 Average 3520 4200 3520 4190 3530 4170 Stdev 1810 2860 1820 2840 1820 2830 p (t-test) 0.010 0.012 0.016 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 159 168 156 171 154 173 UO only 24 48 72 Recovery Period Duration (hr) Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Recovered Cohort Non-recovered Cohort Median 3040 3790 3040 3720 3010 3890 Average 3480 4600 3480 4590 3440 4620 Stdev 2030 2890 2040 2860 2030 2840 p (t-test) 6.4E-5 6.5E-5 1.9E-5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 215 110 212 113 209 116 Recovery Period 24 48 72 Duration (hr) sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.58 0.56 0.62 0.59 0.56 0.62 0.59 0.55 0.63 SE 0.032 0.032 0.034 0.032 0.032 0.033 0.032 0.032 0.033 p Value 0.0081 0.071 3.9E−4 0.0063 0.072 2.7E−4 0.0052 0.094 4.6E−5 nCohort Recovered 187 159 215 184 156 212 183 154 209 nCohort Non- 140 168 110 143 171 113 144 173 116 recovered Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 81% 79% 85% 82% 79% 85% 82% 79% 85% Specificity 30% 30% 30% 30% 29% 30% 31% 29% 31% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 55% 52% 57% 55% 52% 58% 56% 52% 59% Specificity 53% 52% 53% 54% 52% 54% 54% 52% 55% Cutoff Quartile 4 4720 4720 4720 4720 4720 4720 4720 4720 4720 Sensitivity 31% 29% 32% 31% 29% 32% 31% 29% 34% Specificity 79% 79% 78% 79% 79% 78% 79% 79% 79% OR Quartile 2 1.87 1.59 2.32 1.97 1.57 2.44 2.00 1.52 2.57 p Value 0.020 0.070 0.0055 0.012 0.080 0.0032 0.010 0.10 0.0018 Lower limit of 95% CI 1.10 0.963 1.28 1.16 0.948 1.35 1.18 0.918 1.42 Upper limit of 95% CI 3.18 2.64 4.20 3.34 2.59 4.42 3.39 2.51 4.65 OR Quartile 3 1.40 1.20 1.54 1.44 1.17 1.58 1.47 1.17 1.70 p Value 0.13 0.41 0.067 0.10 0.47 0.053 0.084 0.47 0.023 Lower limit of 95% CI 0.905 0.778 0.970 0.927 0.759 0.994 0.950 0.759 1.07 Upper limit of 95% CI 2.18 1.85 2.45 2.23 1.81 2.50 2.28 1.81 2.69 OR Quartile 4 1.68 1.57 1.67 1.71 1.60 1.69 1.68 1.55 1.96 p Value 0.043 0.081 0.052 0.037 0.070 0.046 0.044 0.092 0.010 Lower limit of 95% CI 1.02 0.946 0.996 1.03 0.962 1.01 1.01 0.931 1.17 Upper limit of 95% CI 2.78 2.61 2.79 2.83 2.67 2.82 2.78 2.58 3.26

Example 15. Use of Metalloproteinase Inhibitor 4 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 4 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 15.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.5 34.6 12.5 33.4 12.5 33.4 Average 27.4 227 26.4 224 26.5 221 Stdev 47.8 945 46.5 936 47.2 928 p (t-test) 0.029 0.032 0.038 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 108 209 104 213 100 217 sCr only Median 13.3 34.9 13.3 34.9 13.3 34.9 Average 29.0 246 29.0 244 29.0 241 Stdev 46.7 989 47.0 984 47.3 979 p (t-test) 0.014 0.016 0.017 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 126 190 124 192 122 194 UO only Median 18.5 39.3 18.1 39.3 18.1 36.7 Average 99.9 304 101 275 104 255 Stdev 623 1040 643 976 658 936 p (t-test) 0.033 0.059 0.094 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 222 93 208 107 198 117 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.69 0.67 0.65 0.69 0.67 0.66 0.69 0.67 0.64 SE 0.030 0.030 0.035 0.030 0.030 0.033 0.030 0.030 0.033 p Value 4.8E−10 3.2E−8 2.2E−5 4.2E−10 3.7E−8 3.5E−6 6.4E−10 2.9E−8 3.6E−5 nCohort Recovered 108 126 222 104 124 208 100 122 198 nCohort Non-recovered 209 190 93 213 192 107 217 194 117 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 82% 81% 86% 82% 81% 86% 82% 81% 84% Specificity 39% 33% 30% 39% 34% 31% 40% 34% 30% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 58% 58% 65% 58% 58% 64% 58% 58% 61% Specificity 66% 63% 56% 66% 63% 57% 67% 63% 56% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 33% 35% 39% 33% 35% 37% 33% 35% 36% Specificity 91% 90% 81% 91% 90% 81% 91% 90% 81% OR Quartile 2 2.96 2.07 2.60 3.00 2.15 2.73 3.04 2.23 2.24 p Value 5.2E−5 0.0058 0.0041 4.5E−5 0.0038 0.0015 3.8E−5 0.0024 0.0061 Lower limit of 95% CI 1.75 1.23 1.36 1.77 1.28 1.47 1.79 1.33 1.26 Upper limit of 95% CI 5.00 3.46 5.00 5.08 3.60 5.07 5.17 3.74 3.99 OR Quartile 3 2.69 2.36 2.30 2.75 2.37 2.29 2.81 2.39 1.97 p Value 6.0E−5 2.7E−4 0.0011 5.2E−5 2.6E−4 7.3E−4 4.5E−5 2.5E−4 0.0043 Lower limit of 95% CI 1.66 1.49 1.39 1.68 1.49 1.41 1.71 1.50 1.24 Upper limit of 95% CI 4.36 3.75 3.80 4.48 3.77 3.69 4.62 3.80 3.14 OR Quartile 4 4.94 5.17 2.63 5.28 5.00 2.59 4.92 4.84 2.44 p Value 1.1E−5 1.3E−6 3.9E−4 1.1E−5 2.1E−6 3.8E−4 2.5E−5 3.4E−6 7.9E−4 Lower limit of 95% CI 2.42 2.66 1.54 2.52 2.57 1.53 2.34 2.49 1.45 Upper limit of 95% CI 10.1 10.1 4.48 11.1 9.73 4.37 10.3 9.41 4.10

TABLE 15.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 13.3 36.5 13.3 33.4 13.3 33.4 Average 29.4 257 29.6 254 28.6 249 Stdev 47.4 1010 47.7 1010 46.7 994 p (t-test) 0.0093 0.010 0.012 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 136 181 134 183 129 188 sCr only Median 15.4 36.9 15.9 36.7 15.9 36.7 Average 30.5 274 30.7 272 30.6 269 Stdev 47.1 1050 47.2 1050 47.4 1040 p (t-test) 0.0050 0.0054 0.0060 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 149 168 148 169 146 171 UO only Median 18.2 39.3 18.2 38.8 18.1 33.4 Average 107 297 108 282 111 254 Stdev 625 1060 633 1030 653 964 p (t-test) 0.050 0.067 0.12 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 226 89 220 95 207 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.68 0.67 0.65 0.68 0.67 0.64 0.68 0.67 0.63 SE 0.030 0.030 0.036 0.030 0.030 0.035 0.030 0.030 0.034 p Value 1.0E−9 1.6E−8 4.7E−5 1.5E−9 3.0E−8 7.1E−5 7.1E−10 2.7E−8 1.7E−4 nCohort Recovered 136 149 226 134 148 220 129 146 207 nCohort Non-recovered 181 168 89 183 169 95 188 171 108 Cutoff Quartile 2 8.33 8.33 8.36 8.33 8.33 8.36 8.33 8.33 8.36 Sensitivity 83% 82% 88% 84% 82% 87% 84% 82% 86% Specificity 36% 32% 30% 37% 32% 30% 37% 33% 31% Cutoff Quartile 3 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 Sensitivity 60% 60% 65% 60% 59% 63% 60% 59% 61% Specificity 62% 60% 56% 63% 60% 55% 64% 60% 56% Cutoff Quartile 4 60.9 60.9 61.8 60.9 60.9 61.8 60.9 60.9 61.8 Sensitivity 36% 38% 36% 36% 38% 35% 36% 37% 33% Specificity 90% 89% 79% 90% 89% 79% 90% 89% 79% OR Quartile 2 2.83 2.10 3.05 2.94 2.14 3.03 3.00 2.21 2.77 p Value 1.0E−4 0.0051 0.0016 5.9E−5 0.0042 0.0012 4.1E−5 0.0028 0.0013 Lower limit of 95% CI 1.68 1.25 1.53 1.74 1.27 1.55 1.77 1.31 1.49 Upper limit of 95% CI 4.79 3.53 6.10 4.98 3.59 5.92 5.07 3.72 5.16 OR Quartile 3 2.47 2.24 2.36 2.47 2.19 2.13 2.57 2.19 1.96 p Value 1.1E−4 4.4E−4 9.6E−4 1.1E−4 6.5E−4 0.0027 6.2E−5 6.5E−4 0.0053 Lower limit of 95% CI 1.56 1.43 1.42 1.57 1.39 1.30 1.62 1.40 1.22 Upper limit of 95% CI 3.89 3.52 3.92 3.91 3.43 3.50 4.08 3.43 3.16 OR Quartile 4 5.00 5.12 2.14 4.84 5.03 2.01 4.94 4.86 1.91 p Value 5.6E−7 1.2E−7 0.0057 9.6E−7 1.7E−7 0.010 1.3E−6 3.0E−7 0.015 Lower limit of 95% CI 2.66 2.79 1.25 2.57 2.75 1.18 2.59 2.65 1.13 Upper limit of 95% CI 9.39 9.37 3.67 9.08 9.21 3.43 9.43 8.90 3.22

TABLE 15.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 13.0 38.8 13.0 37.0 13.0 37.0 Average 31.8 280 32.0 277 31.4 271 Stdev 53.3 1060 53.6 1060 52.9 1050 p (t-test) 0.0041 0.0047 0.0056 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 154 163 152 165 148 169 sCr only Median 14.3 38.2 14.5 37.0 14.7 37.0 Average 32.2 290 32.4 289 32.5 287 Stdev 52.3 1090 52.4 1080 52.6 1080 p (t-test) 0.0028 0.0030 0.0032 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 161 156 160 157 159 158 UO only Median 18.1 38.8 18.2 37.8 18.1 33.4 Average 109 286 110 283 112 259 Stdev 628 1050 629 1040 645 986 p (t-test) 0.066 0.071 0.11 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 224 91 223 92 212 103 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.69 0.68 0.63 0.69 0.68 0.62 0.69 0.68 0.62 SE 0.029 0.030 0.036 0.029 0.030 0.036 0.029 0.030 0.034 p Value 3.0E−11 7.2E−10 3.1E−4 9.6E−11 1.5E−9 5.7E−4 1.0E−10 2.7E−9 6.4E−4 nCohort Recovered 154 161 224 152 160 223 148 159 212 nCohort Non-recovered 163 156 91 165 157 92 169 158 103 Cutoff Quartile 2 8.33 8.33 8.36 8.33 8.33 8.36 8.33 8.33 8.36 Sensitivity 85% 83% 87% 85% 83% 86% 85% 84% 84% Specificity 35% 33% 30% 36% 33% 30% 36% 33% 30% Cutoff Quartile 3 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 Sensitivity 63% 62% 64% 62% 62% 63% 62% 61% 62% Specificity 64% 61% 55% 63% 61% 55% 64% 61% 56% Cutoff Quartile 4 60.9 60.9 61.8 60.9 60.9 61.8 60.9 60.9 61.8 Sensitivity 37% 38% 33% 37% 38% 33% 37% 38% 32% Specificity 88% 88% 78% 88% 88% 78% 88% 87% 78% OR Quartile 2 2.98 2.45 2.81 3.09 2.50 2.55 3.07 2.54 2.30 p Value 7.2E−5 9.9E−4 0.0026 4.3E−5 7.9E−4 0.0049 4.2E−5 6.3E−4 0.0074 Lower limit of 95% CI 1.74 1.44 1.44 1.80 1.46 1.33 1.79 1.49 1.25 Upper limit of 95% CI 5.11 4.19 5.50 5.29 4.26 4.91 5.25 4.33 4.23 OR Quartile 3 3.00 2.63 2.18 2.85 2.56 2.10 2.86 2.49 2.06 p Value 2.4E−6 3.0E−5 0.0024 6.8E−6 4.8E−5 0.0036 6.7E−6 7.7E−5 0.0033 Lower limit of 95% CI 1.90 1.67 1.32 1.81 1.63 1.27 1.81 1.58 1.27 Upper limit of 95% CI 4.75 4.13 3.60 4.49 4.02 3.46 4.51 3.91 3.33 OR Quartile 4 4.25 4.41 1.76 4.11 4.33 1.72 4.18 4.26 1.70 p Value 8.4E−7 3.2E−7 0.041 1.5E−6 4.3E−7 0.049 1.5E−6 5.9E−7 0.048 Lower limit of 95% CI 2.39 2.50 1.02 2.31 2.45 1.00 2.33 2.41 1.00 Upper limit of 95% CI 7.56 7.78 3.01 7.30 7.64 2.95 7.50 7.51 2.88

TABLE 15.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 15.1 38.8 15.4 37.0 15.4 36.9 Average 36.0 298 36.4 293 35.9 287 Stdev 62.4 1110 62.9 1100 62.7 1090 p (t-test) 0.0024 0.0030 0.0037 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 453 9030 453 9030 453 9030 n (Patient) 168 149 165 152 161 156 sCr only Median 15.9 37.0 16.3 37.0 16.3 37.0 Average 36.4 305 36.6 303 36.7 301 Stdev 61.8 1130 61.9 1120 62.1 1120 p (t-test) 0.0019 0.0021 0.0023 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 453 9030 453 9030 453 9030 n (Patient) 172 145 171 146 170 147 UO only Median 18.2 39.1 18.1 38.8 18.1 36.7 Average 108 286 109 283 111 265 Stdev 630 1040 631 1040 642 995 p (t-test) 0.065 0.069 0.10 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 223 92 222 93 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.68 0.67 0.64 0.67 0.67 0.64 0.67 0.66 0.63 SE 0.030 0.031 0.035 0.030 0.031 0.035 0.030 0.031 0.035 p Value 2.6E−9 3.2E−8 1.1E−4 6.9E−9 5.9E−8 1.2E−4 8.2E−9 1.0E−7 1.9E−4 nCohort Recovered 168 172 223 165 171 222 161 170 214 nCohort Non-recovered 149 145 92 152 146 93 156 147 101 Cutoff Quartile 2 8.33 8.33 8.36 8.33 8.33 8.36 8.33 8.33 8.36 Sensitivity 84% 83% 86% 84% 83% 86% 84% 83% 85% Specificity 33% 31% 30% 33% 32% 30% 34% 32% 30% Cutoff Quartile 3 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 Sensitivity 63% 62% 64% 62% 62% 65% 62% 61% 63% Specificity 61% 60% 56% 61% 60% 56% 61% 59% 56% Cutoff Quartile 4 60.9 60.9 61.8 60.9 60.9 61.8 60.9 60.9 61.8 Sensitivity 38% 38% 35% 37% 38% 34% 37% 37% 34% Specificity 86% 85% 79% 85% 85% 79% 86% 85% 79% OR Quartile 2 2.54 2.20 2.55 2.67 2.23 2.60 2.64 2.27 2.45 p Value 7.8E−4 0.0041 0.0049 4.9E−4 0.0034 0.0041 4.0E−4 0.0028 0.0048 Lower limit of 95% CI 1.47 1.28 1.33 1.55 1.30 1.36 1.54 1.33 1.31 Upper limit of 95% CI 4.36 3.76 4.91 4.59 3.83 5.00 4.53 3.89 4.55 OR Quartile 3 2.71 2.44 2.24 2.63 2.38 2.30 2.63 2.31 2.21 p Value 1.8E−5 1.1E−4 0.0016 3.0E−5 1.8E−4 0.0011 3.0E−5 2.8E−4 0.0014 Lower limit of 95% CI 1.72 1.55 1.36 1.67 1.51 1.39 1.67 1.47 1.36 Upper limit of 95% CI 4.27 3.84 3.70 4.14 3.73 3.80 4.13 3.63 3.59 OR Quartile 4 3.61 3.59 2.00 3.43 3.53 1.95 3.45 3.47 1.91 p Value 3.8E−6 3.5E−6 0.011 9.1E−6 4.8E−6 0.014 9.5E−6 6.4E−6 0.017 Lower limit of 95% CI 2.10 2.09 1.17 1.99 2.06 1.14 2.00 2.02 1.12 Upper limit of 95% CI 6.23 6.17 3.41 5.90 6.06 3.34 5.98 5.95 3.23

TABLE 15.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 18.1 36.9 18.1 36.3 18.1 35.9 Average 98.8 267 99.6 263 100 261 Stdev 646 951 650 943 651 939 p (t-test) 0.063 0.070 0.074 Min 0.00110 0.00169 0.00110 0.00169 0.00110 0.00169 Max 9030 7570 9030 7570 9030 7570 n (Patient) 203 114 201 116 200 117 sCr only Median 17.0 36.9 17.0 36.9 17.0 36.9 Average 41.1 325 41.1 325 41.1 325 Stdev 69.2 1180 69.2 1180 69.2 1180 p (t-test) 0.0012 0.0012 0.0012 Min 0.00110 0.00169 0.00110 0.00169 0.00110 0.00169 Max 453 9030 453 9030 453 9030 n (Patient) 185 132 185 132 185 132 UO only Median 18.8 37.9 18.5 37.0 19.1 36.7 Average 103 287 103 284 104 279 Stdev 634 1010 635 1010 638 999 p (t-test) 0.051 0.054 0.061 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 217 98 216 99 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.65 0.63 0.63 0.65 0.63 0.62 0.65 0.63 SE 0.033 0.032 0.035 0.033 0.032 0.035 0.033 0.032 0.035 p Value 1.1E−4 2.2E−6 1.5E−4 1.4E−4 2.2E−6 1.5E−4 1.9E−4 2.2E−6 2.8E−4 nCohort 203 185 217 201 185 216 200 185 214 Recovered nCohort 114 132 98 116 132 99 117 132 101 Non- recovered Cutoff 8.33 8.33 8.36 8.33 8.33 8.36 8.33 8.33 8.36 Quartile 2 Sensitivity 82% 83% 85% 83% 83% 85% 83% 83% 85% Specificity 29% 30% 29% 29% 30% 30% 30% 30% 30% Cutoff 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 24.1 Quartile 3 Sensitivity 63% 62% 63% 63% 62% 64% 62% 62% 62% Specificity 57% 58% 56% 57% 58% 56% 57% 58% 56% Cutoff 60.9 60.9 61.8 60.9 60.9 61.8 60.9 60.9 61.8 Quartile 4 Sensitivity 36% 38% 35% 35% 38% 34% 35% 38% 34% Specificity 81% 84% 79% 81% 84% 79% 80% 84% 79% OR 1.93 2.06 2.31 1.99 2.06 2.36 2.03 2.06 2.45 Quartile 2 p Value 0.024 0.0099 0.0082 0.017 0.0099 0.0069 0.015 0.0099 0.0048 Lower limit 1.09 1.19 1.24 1.13 1.19 1.27 1.15 1.19 1.31 of 95% CI Upper limit 3.40 3.56 4.31 3.52 3.56 4.39 3.59 3.56 4.55 of 95% CI OR Quartile 2.29 2.30 2.17 2.27 2.30 2.23 2.20 2.30 2.08 3 p Value 5.9E−4 3.6E−4 0.0019 6.1E−4 3.6E−4 0.0013 9.5E−4 3.6E−4 0.0031 Lower limit 1.43 1.46 1.33 1.42 1.46 1.37 1.38 1.46 1.28 of 95% CI Upper limit 3.66 3.63 3.54 3.63 3.63 3.64 3.51 3.63 3.37 of 95% CI OR Quartile 2.36 3.15 2.03 2.27 3.15 1.99 2.23 3.15 1.91 4 p Value 0.0011 1.9E−5 0.0088 0.0019 1.9E−5 0.011 0.0024 1.9E−5 0.017 Lower limit 1.41 1.86 1.20 1.35 1.86 1.17 1.33 1.86 1.12 of 95% CI Upper limit 3.96 5.33 3.45 3.81 5.33 3.37 3.73 5.33 3.23 of 95% CI

TABLE 15.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2870 3520 2850 3520 2870 3510 Average 3160 4200 3150 4200 3170 4170 Stdev 1580 2680 1550 2670 1570 2660 p (t-test) 2.7E−4 2.6E−4 6.2E−4 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 9100 19700 n (Patient) 105 222 102 225 98 229 sCr only Median 2950 3510 2920 3510 2870 3510 Average 3290 4200 3260 4210 3200 4230 Stdev 1660 2730 1640 2720 1540 2740 p (t-test) 0.0011 5.7E−4 1.9E−4 Min 1080 863 1080 863 1080 863 Max 9820 19700 9820 19700 9100 19700 n (Patient) 121 205 119 207 117 209 UO only Median 3020 3940 3000 3940 3030 3910 Average 3470 4770 3410 4710 3410 4610 Stdev 1940 3070 1910 2980 1910 2920 p (t-test) 5.8E−6 3.0E−6 1.1E−5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 226 99 213 112 202 123 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.61 0.64 0.63 0.62 0.64 0.63 0.62 0.63 SE 0.032 0.032 0.034 0.032 0.031 0.033 0.032 0.031 0.032 p Value 2.8E−5 7.4E−4 5.9E−5 2.3E−5 2.4E−4 1.3E−5 9.2E−5 8.3E−5 5.6E−5 nCohort 105 121 226 102 119 213 98 117 202 Recovered nCohort 222 205 99 225 207 112 229 209 123 Non- recovered Cutoff 2320 2320 2330 2320 2320 2330 2320 2320 2330 Quartile 2 Sensitivity 82% 81% 85% 81% 81% 86% 81% 81% 84% Specificity 39% 36% 29% 39% 36% 31% 39% 37% 30% Cutoff 3280 3280 3280 3280 3280 3280 3280 3280 3280 Quartile 3 Sensitivity 55% 55% 65% 56% 55% 64% 55% 56% 61% Specificity 61% 58% 56% 62% 59% 57% 61% 60% 56% Cutoff 4720 4710 4720 4720 4710 4720 4720 4710 4720 Quartile 4 Sensitivity 30% 31% 35% 30% 31% 35% 30% 31% 34% Specificity 86% 84% 79% 86% 85% 80% 86% 85% 80% OR Quartile 2.83 2.35 2.31 2.81 2.44 2.64 2.66 2.53 2.23 2 p Value 8.4E−5 0.0010 0.0081 9.8E−5 6.4E−4 0.0017 2.4E−4 3.8E−4 0.0055 Lower limit 1.68 1.41 1.24 1.67 1.46 1.44 1.58 1.52 1.27 of 95% CI Upper limit 4.75 3.91 4.29 4.73 4.06 4.82 4.49 4.23 3.92 of 95% CI OR Quartile 1.94 1.65 2.35 2.02 1.75 2.41 1.93 1.86 2.02 3 p Value 0.0061 0.030 6.3E−4 0.0040 0.016 2.6E−4 0.0075 0.0082 0.0025 Lower limit 1.21 1.05 1.44 1.25 1.11 1.50 1.19 1.17 1.28 of 95% CI Upper limit 3.11 2.60 3.82 3.26 2.76 3.87 3.13 2.94 3.20 of 95% CI OR Quartile 2.59 2.38 2.08 2.72 2.51 2.11 2.53 2.66 2.10 4 p Value 0.0025 0.0030 0.0059 0.0019 0.0019 0.0043 0.0040 0.0012 0.0042 Lower limit 1.40 1.34 1.24 1.45 1.40 1.26 1.35 1.47 1.26 of 95% CI Upper limit 4.81 4.22 3.51 5.12 4.49 3.53 4.77 4.80 3.49 of 95% CI

TABLE 15.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2930 3560 2920 3630 2980 3520 Average 3230 4310 3210 4320 3250 4270 Stdev 1540 2810 1520 2800 1530 2790 p (t-test) 6.5E−5 3.8E−5 1.7E−4 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 9100 19700 n (Patient) 134 193 133 194 128 199 sCr only Median 2980 3520 2950 3540 2920 3540 Average 3310 4320 3290 4330 3240 4360 Stdev 1610 2850 1590 2850 1510 2860 p (t-test) 1.5E−4 8.7E−5 2.7E−5 Min 1080 863 1080 863 1080 863 Max 9820 19700 9820 19700 9100 19700 n (Patient) 146 181 145 182 143 184 UO only Median 3060 3890 3040 3930 3050 3920 Average 3530 4670 3500 4670 3460 4600 Stdev 2060 2970 2060 2910 2040 2840 p (t-test) 9.7E−5 5.0E−5 3.9E−5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 231 94 225 100 210 115 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.62 0.61 0.62 0.63 0.62 0.63 0.61 0.62 0.63 SE 0.031 0.031 0.035 0.031 0.031 0.034 0.031 0.031 0.033 p Value 6.4E−5 2.9E−4 4.3E−4 2.8E−5 1.4E−4 1.1E−4 2.5E−4 5.3E−5 8.3E−5 nCohort 134 146 231 133 145 225 128 143 210 Recovered nCohort 193 181 94 194 182 100 199 184 115 Non- recovered Cutoff 2320 2320 2330 2320 2320 2330 2320 2320 2330 Quartile 2 Sensitivity 82% 82% 86% 82% 82% 87% 81% 82% 84% Specificity 35% 34% 29% 35% 34% 30% 34% 34% 30% Cutoff 3280 3280 3280 3280 3280 3280 3280 3280 3280 Quartile 3 Sensitivity 56% 56% 62% 56% 56% 62% 55% 57% 60% Specificity 58% 57% 55% 59% 57% 55% 58% 58% 55% Cutoff 4720 4720 4720 4720 4720 4720 4720 4720 4720 Quartile 4 Sensitivity 32% 32% 33% 32% 32% 34% 31% 33% 35% Specificity 84% 84% 78% 85% 84% 79% 84% 85% 80% OR 2.44 2.27 2.60 2.48 2.30 2.90 2.22 2.39 2.31 Quartile 2 p Value 6.1E−4 0.0017 0.0040 4.8E−4 0.0013 0.0013 0.0021 8.6E−4 0.0049 Lower limit 1.46 1.36 1.36 1.49 1.38 1.52 1.34 1.43 1.29 of 95% CI Upper limit 4.06 3.77 4.98 4.14 3.84 5.54 3.69 3.98 4.14 of 95% CI OR 1.77 1.66 1.93 1.82 1.71 2.00 1.69 1.80 1.85 Quartile 3 p Value 0.012 0.023 0.0084 0.0087 0.017 0.0047 0.021 0.0092 0.0089 Lower limit 1.13 1.07 1.18 1.16 1.10 1.24 1.08 1.16 1.17 of 95% CI Upper limit 2.76 2.58 3.16 2.84 2.65 3.24 2.65 2.80 2.94 of 95% CI OR 2.49 2.40 1.74 2.65 2.54 1.90 2.44 2.66 2.13 Quartile 4 p Value 0.0013 0.0014 0.041 6.8E−4 7.5E−4 0.016 0.0019 4.7E−4 0.0037 Lower limit 1.43 1.40 1.02 1.51 1.48 1.13 1.39 1.54 1.28 of 95% CI Upper limit 4.34 4.10 2.95 4.66 4.38 3.20 4.29 4.61 3.56 of 95% CI

TABLE 15.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3100 3490 3080 3510 3120 3470 Average 3460 4230 3430 4250 3470 4200 Stdev 1830 2810 1810 2800 1820 2790 p (t-test) 0.0042 0.0021 0.0060 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 154 173 152 175 148 179 sCr only Median 3110 3480 3100 3490 3090 3500 Average 3500 4230 3480 4240 3440 4280 Stdev 1870 2830 1860 2820 1790 2850 p (t-test) 0.0060 0.0042 0.0017 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 161 166 160 167 159 168 UO only Median 3080 3720 3050 3720 3070 3720 Average 3540 4640 3520 4670 3490 4580 Stdev 2050 2980 2040 2970 2040 2890 p (t-test) 1.7E−4 6.3E−5 1.1E−4 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 230 95 228 97 215 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.58 0.57 0.61 0.59 0.58 0.62 0.57 0.58 0.62 SE 0.031 0.032 0.035 0.031 0.031 0.035 0.032 0.031 0.034 p Value 0.015 0.020 0.0012 0.0062 0.013 3.8E−4 0.022 0.0075 4.7E−4 nCohort 154 161 230 152 160 228 148 159 215 Recovered nCohort 173 166 95 175 167 97 179 168 110 Non- recovered Cutoff 2320 2320 2330 2320 2320 2330 2320 2320 2330 Quartile 2 Sensitivity 80% 81% 85% 81% 81% 86% 80% 81% 85% Specificity 31% 31% 29% 32% 31% 29% 31% 31% 30% Cutoff 3280 3280 3280 3280 3280 3280 3280 3280 3280 Quartile 3 Sensitivity 53% 53% 59% 54% 53% 60% 53% 54% 58% Specificity 53% 53% 53% 54% 53% 54% 53% 53% 54% Cutoff 4720 4720 4720 4720 4720 4720 4720 4720 4720 Quartile 4 Sensitivity 29% 30% 32% 30% 30% 33% 29% 30% 33% Specificity 79% 80% 77% 80% 80% 78% 80% 81% 79% OR 1.85 1.89 2.38 1.91 1.92 2.47 1.79 1.95 2.32 Quartile 2 p Value 0.017 0.015 0.0075 0.012 0.012 0.0052 0.024 0.010 0.0055 Lower limit 1.12 1.13 1.26 1.15 1.15 1.31 1.08 1.17 1.28 of 95% CI Upper limit 3.07 3.14 4.49 3.18 3.19 4.65 2.97 3.25 4.20 of 95% CI OR 1.29 1.26 1.65 1.36 1.29 1.74 1.23 1.33 1.63 Quartile 3 p Value 0.25 0.29 0.042 0.17 0.25 0.024 0.35 0.20 0.039 Lower limit 0.837 0.817 1.02 0.879 0.837 1.08 0.797 0.858 1.02 of 95% CI Upper limit 2.00 1.95 2.68 2.10 2.00 2.82 1.91 2.05 2.59 of 95% CI OR Quartile 1.55 1.62 1.58 1.72 1.71 1.75 1.61 1.80 1.79 4 p Value 0.092 0.061 0.092 0.039 0.039 0.037 0.069 0.024 0.027 Lower limit 0.931 0.978 0.928 1.03 1.03 1.03 0.963 1.08 1.07 of 95% CI Upper limit 2.58 2.70 2.69 2.88 2.85 2.97 2.69 3.00 2.99 of 95% CI

TABLE 15.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3080 3510 3040 3520 3090 3500 Average 3460 4290 3430 4310 3460 4250 Stdev 1790 2890 1780 2870 1790 2850 p (t-test) 0.0020 9.9E−4 0.0030 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 166 161 163 164 159 168 sCr only Median 3110 3500 3100 3510 3090 3510 Average 3510 4250 3490 4270 3460 4300 Stdev 1850 2880 1840 2880 1780 2900 p (t-test) 0.0056 0.0038 0.0015 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 169 158 168 159 167 160 UO only Median 3050 3860 3040 3790 3070 3720 Average 3500 4690 3500 4680 3500 4590 Stdev 2020 2980 2030 2960 2040 2910 p (t-test) 3.3E−5 3.6E−5 1.1E−4 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 226 99 225 100 217 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.58 0.57 0.63 0.59 0.57 0.63 0.58 0.58 0.62 SE 0.032 0.032 0.035 0.031 0.032 0.034 0.032 0.032 0.034 p Value 0.013 0.031 3.0E−4 0.0046 0.021 2.7E−4 0.017 0.012 6.2E−4 nCohort Recovered 166 169 226 163 168 225 159 167 217 nCohort Non-recovered 161 158 99 164 159 100 168 160 108 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 80% 80% 86% 80% 80% 86% 80% 80% 85% Specificity 30% 30% 30% 31% 30% 30% 30% 30% 30% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 54% 53% 59% 55% 53% 59% 54% 54% 57% Specificity 54% 53% 54% 55% 53% 54% 53% 53% 53% Cutoff Quartile 4 4720 4720 4720 4720 4720 4720 4720 4720 4720 Sensitivity 29% 29% 33% 30% 30% 33% 29% 30% 32% Specificity 79% 79% 78% 80% 79% 78% 79% 80% 78% OR Quartile 2 1.74 1.65 2.56 1.83 1.68 2.60 1.70 1.71 2.46 p Value 0.034 0.053 0.0036 0.021 0.046 0.0030 0.039 0.039 0.0036 Lower limit of 95% CI 1.04 0.994 1.36 1.10 1.01 1.38 1.03 1.03 1.34 Upper limit of 95% CI 2.89 2.75 4.82 3.04 2.80 4.91 2.83 2.85 4.50 OR Quartile 3 1.36 1.26 1.63 1.46 1.29 1.67 1.33 1.33 1.55 p Value 0.17 0.29 0.045 0.087 0.25 0.034 0.20 0.20 0.066 Lower limit of 95% CI 0.880 0.818 1.01 0.946 0.838 1.04 0.858 0.859 0.972 Upper limit of 95% CI 2.10 1.95 2.63 2.26 2.00 2.70 2.05 2.05 2.47 OR Quartile 4 1.54 1.52 1.81 1.68 1.59 1.77 1.57 1.68 1.73 p Value 0.092 0.10 0.027 0.046 0.070 0.033 0.081 0.045 0.037 Lower limit of 95% CI 0.932 0.917 1.07 1.01 0.963 1.05 0.946 1.01 1.03 Upper limit of 95% CI 2.56 2.51 3.05 2.79 2.64 2.99 2.61 2.78 2.91

TABLE 15.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3100 3510 3080 3520 3090 3510 Average 3560 4390 3550 4390 3550 4380 Stdev 2090 2830 2100 2810 2100 2800 p (t-test) 0.0026 0.0020 0.0025 Min 1080 863 1080 863 1080 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 204 123 202 125 201 126 sCr only Median 3080 3510 3080 3510 3080 3510 Average 3480 4370 3480 4370 3480 4370 Stdev 1770 3010 1770 3010 1770 3010 p (t-test) 8.8E−4 8.8E−4 8.8E−4 Min 1080 863 1080 863 1080 863 Max 14500 19700 14500 19700 14500 19700 n (Patient) 184 143 184 143 184 143 UO only Median 3040 3860 3040 3790 3040 3790 Average 3470 4670 3470 4660 3470 4650 Stdev 2030 2910 2030 2900 2030 2880 p (t-test) 2.3E−5 2.5E−5 2.3E−5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 220 105 219 106 217 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.58 0.58 0.63 0.59 0.58 0.63 0.59 0.58 0.63 SE 0.033 0.032 0.034 0.033 0.032 0.034 0.033 0.032 0.034 p Value 0.011 0.013 1.2E−4 0.0064 0.013 1.1E−4 0.0079 0.013 8.0E−5 nCohort Recovered 204 184 220 202 184 219 201 184 217 nCohort Non-recovered 123 143 105 125 143 106 126 143 108 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 83% 82% 87% 83% 82% 87% 83% 82% 87% Specificity 30% 30% 30% 30% 30% 31% 30% 30% 31% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 55% 55% 58% 56% 55% 58% 56% 55% 58% Specificity 53% 53% 54% 53% 53% 54% 53% 53% 54% Cutoff Quartile 4 4720 4720 4720 4720 4720 4720 4720 4720 4720 Sensitivity 29% 30% 32% 30% 30% 32% 29% 30% 32% Specificity 77% 79% 78% 78% 79% 78% 78% 79% 78% OR Quartile 2 2.07 1.97 2.85 2.14 1.97 2.90 2.18 1.97 3.00 p Value 0.010 0.012 0.0012 0.0073 0.012 9.6E−4 0.0061 0.012 6.5E−4 Lower limit of 95% CI 1.19 1.16 1.51 1.23 1.16 1.54 1.25 1.16 1.60 Upper limit of 95% CI 3.62 3.34 5.35 3.74 3.34 5.45 3.80 3.34 5.64 OR Quartile 3 1.39 1.37 1.60 1.46 1.37 1.65 1.42 1.37 1.64 p Value 0.15 0.16 0.049 0.097 0.16 0.037 0.12 0.16 0.038 Lower limit of 95% CI 0.887 0.882 1.00 0.934 0.882 1.03 0.910 0.882 1.03 Upper limit of 95% CI 2.18 2.12 2.57 2.29 2.12 2.63 2.23 2.12 2.61 OR Quartile 4 1.42 1.60 1.72 1.47 1.60 1.68 1.44 1.60 1.73 p Value 0.18 0.067 0.041 0.14 0.067 0.049 0.16 0.067 0.037 Lower limit of 95% CI 0.855 0.967 1.02 0.883 0.967 1.00 0.868 0.967 1.03 Upper limit of 95% CI 2.36 2.64 2.88 2.44 2.64 2.83 2.39 2.64 2.91

Example 16. Use of Metalloproteinase Inhibitor 4 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 4 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 16.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 16.3 51.9 16.5 61.5 17.0 56.6 Average 37.9 382 37.4 440 78.7 394 Stdev 62.5 1270 60.8 1370 589 1120 p (t-test) 1.3E−4 1.6E−5 0.0014 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 9030 7570 n (Patient) 205 112 221 96 236 81 sCr only Median 16.9 48.7 16.9 56.6 18.2 53.0 Average 38.8 392 38.9 449 80.3 389 Stdev 62.9 1290 61.8 1390 590 1130 p (t-test) 1.0E−4 1.4E−5 0.0019 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 9030 7570 n (Patient) 208 108 223 93 235 81 UO only Median 20.5 54.3 20.7 55.6 20.8 57.9 Average 99.1 580 98.5 624 98.3 640 Stdev 568 1540 565 1590 564 1610 p (t-test) 2.2E−4 9.2E−5 6.7E−5 Min 0.00110 1.76 0.00110 1.76 0.00110 1.76 Max 9030 7570 9030 7570 9030 7570 n (Patient) 275 40 278 37 279 36 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.71 0.69 0.69 0.74 0.72 0.69 0.73 0.70 0.70 SE 0.031 0.032 0.049 0.032 0.033 0.050 0.035 0.036 0.051 p Value 2.1E−11 1.8E−9 8.2E−5 6.9E−14 4.2E−11 1.3E−4 2.0E−11 3.0E−8 1.2E−4 nCohort Non-persistent 205 208 275 221 223 278 236 235 279 nCohort Persistent 112 108 40 96 93 37 81 81 36 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 86% 84% 88% 89% 87% 86% 89% 86% 86% Specificity 31% 30% 27% 31% 30% 27% 30% 29% 27% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 71% 69% 80% 74% 72% 78% 77% 73% 78% Specificity 61% 60% 54% 60% 59% 54% 59% 58% 53% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 46% 45% 45% 50% 48% 46% 48% 46% 47% Specificity 86% 86% 78% 86% 85% 78% 83% 82% 78% OR Quartile 2 2.66 2.27 2.58 3.43 2.90 2.32 3.37 2.59 2.24 p Value 0.0016 0.0070 0.057 4.6E−4 0.0019 0.092 0.0014 0.0073 0.11 Lower limit of 95% CI 1.45 1.25 0.973 1.72 1.48 0.872 1.60 1.29 0.839 Upper limit of 95% CI 4.88 4.13 6.83 6.85 5.67 6.18 7.12 5.19 5.97 OR Quartile 3 3.74 3.21 4.73 4.29 3.74 4.19 4.68 3.68 4.01 p Value 1.6E−7 3.3E−6 1.7E−4 6.9E−8 8.9E−7 6.0E−4 1.5E−7 3.9E−6 9.0E−4 Lower limit of 95% CI 2.28 1.97 2.10 2.53 2.21 1.85 2.63 2.12 1.77 Upper limit of 95% CI 6.13 5.25 10.6 7.29 6.32 9.48 8.32 6.41 9.11 OR Quartile 4 5.07 4.93 2.87 5.91 5.21 2.96 4.42 3.86 3.13 p Value 4.0E−9 7.8E−9 0.0025 2.1E−10 3.2E−9 0.0026 1.3E−7 1.5E−6 0.0017 Lower limit of 95% CI 2.95 2.87 1.45 3.41 3.02 1.46 2.55 2.23 1.54 Upper limit of 95% CI 8.72 8.47 5.69 10.2 9.00 6.00 7.66 6.70 6.39

TABLE 16.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 16.3 43.9 16.3 56.6 16.5 56.1 Average 35.1 336 34.1 385 78.4 340 Stdev 57.4 1180 55.3 1260 611 1030 p (t-test) 5.9E−4 9.1E−5 0.0052 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 9030 7570 n (Patient) 186 131 204 113 219 98 sCr only Median 16.9 42.2 16.7 51.9 17.0 46.3 Average 37.9 348 36.9 389 81.6 332 Stdev 60.8 1210 59.2 1280 612 1030 P (t-test) 4.7E−4 1.0E−4 0.0075 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 9030 7570 n (Patient) 192 124 206 110 218 98 UO only Median 20.0 43.9 20.4 50.8 20.5 51.9 Average 101 416 100 443 100 451 Stdev 588 1280 584 1330 583 1340 p (t-test) 0.0048 0.0027 0.0023 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 256 59 260 55 261 54 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.70 0.67 0.66 0.73 0.71 0.66 0.73 0.69 0.66 SE 0.030 0.032 0.042 0.031 0.032 0.043 0.032 0.034 0.043 p Value 6.6E−11 3.6E−8 1.0E−4 2.6E−14 8.9E−11 1.4E−4 1.3E−12 2.5E−8 1.4E−4 nCohort Non- persistent 186 192 256 204 206 260 219 218 261 nCohort Persistent 131 124 59 113 110 55 98 98 54 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 85% 83% 86% 87% 85% 85% 88% 86% 85% Specificity 32% 30% 28% 31% 31% 27% 31% 30% 27% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 66% 65% 71% 72% 69% 71% 73% 68% 70% Specificity 61% 59% 55% 62% 60% 54% 60% 58% 54% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 43% 42% 41% 48% 45% 42% 47% 43% 43% Specificity 87% 86% 79% 87% 86% 78% 84% 83% 79% OR Quartile 2 2.58 2.12 2.45 2.99 2.59 2.21 3.16 2.55 2.15 p Value 0.0011 0.0086 0.027 5.3E−4 0.0021 0.052 7.5E−4 0.0039 0.061 Lower limit of 95% CI 1.46 1.21 1.11 1.61 1.41 0.994 1.62 1.35 0.967 Upper limit of 95% CI 4.55 3.72 5.41 5.54 4.75 4.90 6.17 4.81 4.78 OR Quartile 3 3.13 2.66 2.98 4.09 3.38 2.89 4.20 3.02 2.79 p Value 1.7E−6 4.1E−5 5.0E−4 2.8E−8 1.2E−6 9.9E−4 7.8E−8 1.7E−5 0.0015 Lower limit of 95% CI 1.96 1.67 1.61 2.49 2.07 1.54 2.49 1.82 1.48 Upper limit of 95% CI 5.00 4.24 5.51 6.72 5.52 5.43 7.09 4.99 5.25 OR Quartile 4 5.04 4.41 2.51 6.27 5.09 2.62 4.81 3.67 2.72 p Value 8.7E−9 7.5E−8 0.0026 7.7E−11 4.4E−9 0.0021 1.1E−8 1.8E−6 0.0015 Lower limit of 95% CI 2.90 2.57 1.38 3.61 2.95 1.42 2.81 2.15 1.47 Upper limit of 95% CI 8.74 7.58 4.56 10.9 8.76 4.83 8.26 6.26 5.02

TABLE 16.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 15.8 42.0 16.0 53.0 16.3 51.9 Average 33.1 315 32.1 350 79.7 305 Stdev 52.8 1140 51.2 1200 631 967 p (t-test) 0.0012 3.0E−4 0.013 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 9030 7570 n (Patient) 175 142 190 127 205 112 sCr only Median 16.4 42.0 16.3 46.5 17.0 45.0 Average 34.5 329 34.8 360 81.7 309 Stdev 53.5 1170 54.9 1230 626 985 p (t-test) 7.6E−4 2.5E−4 0.013 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 9030 7570 n (Patient) 182 134 195 121 208 108 UO only Median 19.1 40.3 19.6 47.4 19.6 47.4 Average 103 365 101 391 101 391 Stdev 600 1190 594 1240 594 1240 p (t-test) 0.013 0.0075 0.0075 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 246 69 251 64 251 64 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.69 0.68 0.65 0.72 0.70 0.66 0.71 0.68 0.66 SE 0.030 0.031 0.039 0.030 0.031 0.040 0.031 0.033 0.040 p Value 9.5E−11 9.9E−9 1.3E−4 4.7E−14 4.9E−11 1.0E−4 1.8E−11 1.9E−8 1.0E−4 nCohort Non- 175 182 246 190 195 251 205 208 251 persistent nCohort Persistent 142 134 69 127 121 64 112 108 64 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 85% 83% 86% 87% 85% 84% 87% 85% 84% Specificity 33% 31% 28% 33% 31% 27% 31% 30% 27% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 65% 64% 70% 69% 68% 70% 70% 67% 70% Specificity 62% 60% 55% 63% 61% 55% 60% 59% 55% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 41% 41% 39% 45% 44% 41% 44% 42% 41% Specificity 87% 87% 79% 88% 87% 79% 85% 84% 79% OR Quartile 2 2.63 2.14 2.30 3.13 2.60 2.05 2.94 2.50 2.05 p Value 6.0E−4  0.0064 0.024 1.6E−4  0.0013 0.054 6.5E−4  0.0031 0.054 Lower limit of 95% CI 1.51 1.24 1.11 1.73 1.45 0.987 1.58 1.36 0.987 Upper limit of 95% CI 4.58 3.71 4.75 5.68 4.68 4.25 5.45 4.59 4.25 OR Quartile 3 3.13 2.74 2.83 3.78 3.29 2.89 3.51 2.84 2.89 p Value 1.2E−6  1.9E−5 3.6E−4 4.9E−8  1.0E−6  4.3E−4 5.2E−7  2.6E−5 4.3E−4 Lower limit of 95% CI 1.98 1.73 1.60 2.34 2.04 1.60 2.15 1.75 1.60 Upper limit of 95% CI 4.97 4.34 5.00 6.10 5.31 5.22 5.74 4.61 5.22 OR Quartile 4 4.80 4.58 2.40 5.91 5.07 2.56 4.37 3.66 2.56 p Value 3.6E−8  5.9E−8 0.0027 4.6E−10 6.2E−9  0.0016 6.4E−8  1.7E−6 0.0016 Lower limit of 95% CI 2.75 2.64 1.35 3.38 2.93 1.43 2.56 2.15 1.43 Upper limit of 95% CI 8.39 7.95 4.25 10.3 8.76 4.58 7.45 6.21 4.58

TABLE 16.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 15.8 42.0 16.3 51.9 16.3 48.7 Average 32.9 311 32.2 347 36.2 373 Stdev 53.0 1130 51.3 1190 59.1 1250 p (t-test) 0.0013 3.4E−4 1.6E−4 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 173 144 189 128 201 116 sCr only Median 16.7 41.1 16.3 46.3 17.0 45.0 Average 34.8 324 33.8 354 37.6 381 Stdev 53.8 1160 52.5 1210 59.9 1270 p (t-test) 9.2E−4 3.0E−4 1.4E−4 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 180 136 192 124 204 112 UO only Median 18.2 42.1 18.8 47.4 18.8 47.4 Average 104 337 102 363 102 363 Stdev 608 1140 601 1180 601 1180 p (t-test) 0.022 0.013 0.013 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 239 76 245 70 245 70 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.67 0.66 0.72 0.70 0.66 0.71 0.69 0.66 SE 0.030 0.031 0.038 0.030 0.031 0.039 0.031 0.032 0.039 p Value 8.7E−11 3.9E−8 2.5E−5 1.6E−13 6.9E−11 3.7E−5 3.8E−11 6.9E−9 3.7E−5 nCohort Non-persistent 173 180 239 189 192 245 201 204 245 nCohort 144 136 76 128 124 70 116 112 70 Persistent Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 84% 82% 86% 86% 85% 84% 86% 85% 84% Specificity 32% 31% 28% 32% 31% 28% 31% 30% 28% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 65% 63% 70% 69% 67% 70% 68% 66% 70% Specificity 62% 60% 56% 62% 61% 56% 60% 59% 56% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 41% 40% 39% 45% 44% 40% 43% 42% 40% Specificity 88% 87% 79% 88% 87% 79% 85% 84% 79% OR Quartile 2 2.52 2.05 2.35 2.91 2.51 2.06 2.85 2.44 2.06 p Value 9.5E−4  0.0094 0.016 3.4E−4  0.0017 0.043 7.0E−4  0.0034 0.043 Lower limit of 95% CI 1.46 1.19 1.17 1.62 1.41 1.02 1.56 1.34 1.02 Upper limit of 95% CI 4.36 3.53 4.72 5.22 4.47 4.16 5.23 4.43 4.16 OR Quartile 3 3.12 2.58 2.94 3.66 3.16 2.91 3.23 2.78 2.91 p Value 1.3E−6  5.1E−5 1.3E−4 9.2E−8  1.9E−6  2.4E−4 1.9E−6  3.0E−5 2.4E−4 Lower limit of 95% CI 1.97 1.63 1.69 2.27 1.97 1.65 1.99 1.72 1.65 Upper limit of 95% CI 4.95 4.08 5.11 5.88 5.07 5.15 5.23 4.50 5.15 OR Quartile 4 5.02 4.41 2.53 5.79 5.15 2.54 4.32 3.89 2.54 p Value 2.1E−8  1.2E−7 0.0011 6.9E−10 5.2E−9  0.0013 8.1E−8  5.7E−7 0.0013 Lower limit of 95% CI 2.86 2.55 1.45 3.32 2.97 1.44 2.53 2.28 1.44 Upper limit of 95% CI 8.83 7.65 4.41 10.1 8.93 4.48 7.37 6.62 4.48

TABLE 16.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 15.8 41.1 15.8 50.8 16.3 46.3 Average 32.4 304 31.4 336 36.1 362 Stdev 52.7 1110 50.9 1170 59.5 1230 p (t-test) 0.0017 4.8E−4 2.5E−4 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 169 148 184 133 197 120 sCr only Median 16.4 40.2 16.3 45.0 16.7 43.2 Average 34.4 317 33.4 345 37.3 370 Stdev 53.5 1140 52.2 1190 59.8 1250 p (t-test) 0.0012 4.1E−4 2.0E−4 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 176 140 188 128 200 116 UO only Median 18.1 46.1 18.2 48.4 18.2 48.4 Average 97.3 342 96.6 354 96.6 354 Stdev 611 1110 607 1130 607 1130 p (t-test) 0.014 0.011 0.011 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 234 81 237 78 237 78 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.67 0.68 0.72 0.70 0.68 0.71 0.68 0.68 SE 0.030 0.031 0.036 0.029 0.031 0.037 0.031 0.032 0.037 p Value 8.1E−11 3.4E−8 1.2E−6 2.5E−14 6.5E−11 1.6E−6 3.6E−11 7.2E−9 1.6E−6 nCohort Non- 169 176 234 184 188 237 197 200 237 persistent nCohort Persistent 148 140 81 133 128 78 120 116 78 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 84% 82% 86% 86% 84% 86% 86% 84% 86% Specificity 33% 31% 29% 33% 31% 29% 31% 30% 29% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 65% 63% 70% 69% 66% 71% 68% 66% 71% Specificity 63% 60% 57% 64% 61% 57% 61% 59% 57% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 41% 40% 41% 44% 43% 41% 42% 41% 41% Specificity 88% 87% 80% 89% 87% 80% 85% 84% 80% OR Quartile 2 2.49 2.04 2.61 2.90 2.47 2.45 2.78 2.39 2.45 p Value 9.7E−4  0.0096 0.0069 2.8E−4  0.0018 0.012 7.4E−4  0.0036 0.012 Lower limit of 95% CI 1.45 1.19 1.30 1.63 1.40 1.22 1.54 1.33 1.22 Upper limit of 95% CI 4.29 3.49 5.23 5.16 4.36 4.92 5.04 4.29 4.92 OR Quartile 3 3.11 2.56 3.13 3.92 3.11 3.11 3.36 2.73 3.11 p Value 1.3E−6  5.4E−5 3.8E−5 1.7E−8  2.1E−6  5.3E−5 7.1E−7  3.4E−5 5.3E−5 Lower limit of 95% CI 1.96 1.62 1.82 2.44 1.95 1.79 2.08 1.70 1.79 Upper limit of 95% CI 4.92 4.05 5.38 6.30 4.98 5.39 5.43 4.40 5.39 OR Quartile 4 5.08 4.43 2.81 6.19 5.15 2.81 4.28 3.85 2.81 p Value 2.4E−8  1.3E−7 2.2E−4 3.4E−10 6.4E−9  2.5E−4 1.0E−7  6.9E−7 2.5E−4 Lower limit of 95% CI 2.87 2.55 1.62 3.50 2.96 1.62 2.51 2.26 1.62 Upper limit of 95% CI 8.99 7.71 4.86 10.9 8.95 4.89 7.31 6.55 4.89

TABLE 16.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3120 3720 3140 3810 3150 3810 Average 3490 4520 3560 4520 3630 4500 Stdev 1640 3270 1820 3280 2090 3090 p (t-test) 2.0E−4 8.7E−4 0.0039 Min 1080 863 1080 863 1080 863 Max 9820 19700 14500 19700 19700 18800 n (Patient) 206 121 222 105 238 89 sCr only Median 3100 3620 3090 3860 3120 3670 Average 3470 4540 3530 4580 3620 4490 Stdev 1630 3310 1810 3300 2080 3080 p (t-test) 1.1E−4 2.7E−4 0.0035 Min 1080 863 1080 863 1080 863 Max 9820 19700 14500 19700 19700 18800 n (Patient) 208 118 223 103 236 90 UO only Median 3140 4580 3150 4610 3140 4750 Average 3580 5670 3580 5740 3570 5940 Stdev 2020 3670 2010 3740 2010 3720 p (t-test) 5.0E−8 3.4E−8 2.5E−9 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 281 44 283 42 285 40 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.58 0.69 0.57 0.59 0.69 0.57 0.57 0.72 SE 0.033 0.033 0.047 0.034 0.035 0.048 0.036 0.036 0.048 p Value 0.022 0.018 5.4E−5 0.036 0.013 8.5E−5 0.048 0.039 5.3E−6 nCohort Non-persistent 206 208 281 222 223 283 238 236 285 nCohort Persistent 121 118 44 105 103 42 89 90 40 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 79% 79% 91% 80% 80% 90% 80% 79% 95% Specificity 28% 27% 27% 27% 27% 27% 27% 27% 28% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 56% 56% 68% 56% 57% 67% 57% 57% 70% Specificity 53% 53% 53% 53% 53% 52% 53% 53% 53% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 33% 34% 45% 33% 35% 48% 33% 33% 50% Specificity 80% 80% 78% 79% 79% 78% 78% 78% 78% OR Quartile 2 1.47 1.40 3.77 1.52 1.47 3.55 1.45 1.36 7.29 p Value 0.16 0.21 0.014 0.15 0.18 0.019 0.22 0.30 0.0071 Lower limit of 95% CI 0.860 0.821 1.31 0.864 0.838 1.23 0.803 0.760 1.72 Upper limit of 95% CI 2.51 2.40 10.9 2.66 2.58 10.3 2.62 2.44 30.9 OR Quartile 3 1.47 1.45 2.38 1.43 1.53 2.19 1.48 1.45 2.59 p Value 0.094 0.11 0.012 0.13 0.075 0.024 0.11 0.14 0.0090 Lower limit of 95% CI 0.936 0.922 1.21 0.896 0.958 1.11 0.909 0.888 1.27 Upper limit of 95% CI 2.31 2.29 4.69 2.28 2.46 4.34 2.43 2.36 5.30 OR Quartile 4 1.93 2.03 2.94 1.86 2.07 3.24 1.69 1.77 3.60 p Value 0.011 0.0066 0.0013 0.019 0.0061 5.6E−4 0.057 0.037 2.3E−4 Lower limit of 95% CI 1.16 1.22 1.53 1.11 1.23 1.66 0.985 1.04 1.82 Upper limit of 95% CI 3.21 3.37 5.68 3.13 3.47 6.32 2.89 3.02 7.10

TABLE 16.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal staus is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3110 3720 3120 3810 3140 3770 Average 3420 4470 3510 4470 3600 4420 Stdev 1590 3130 1800 3120 2090 2930 p (t-test) 9.5E-5 4.3E-4 0.0035 Min 1080 863 1080 863 1080 863 Max 9820 19700 14500 19700 19700 18800 n (Patient) 187 140 204 123 219 108 sCr only Median 3100 3620 3090 3810 3130 3520 Average 3420 4490 3490 4480 3600 4380 Stdev 1580 3180 1790 3140 2090 2930 p (t-test) 8.1E-5 3.4E-4 0.0059 Min 1080 863 1080 863 1080 863 Max 9820 19700 14500 19700 19700 18800 n (Patient) 192 134 205 121 217 109 UO only Median 3140 3990 3140 4150 3130 4350 Average 3570 5020 3570 5090 3560 5210 Stdev 2040 3300 2030 3360 2030 3360 p (t-test) 1.2E−5 5.8E−6 1.2E−6 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 260 65 263 62 265 60 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.59 0.59 0.64 0.58 0.59 0.64 0.58 0.57 0.66 SE 0.032 0.032 0.040 0.033 0.033 0.041 0.034 0.034 0.041 p Value 0.0062 0.0078 7.5E−4 0.010 0.0084 6.1E−4 0.025 0.039 1.0E−4 nCohort Non-persistent 187 192 260 204 205 263 219 217 265 nCohort Persistent 140 134 65 123 121 62 108 109 60 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 80% 80% 88% 81% 81% 89% 81% 81% 92% Specificity 29% 29% 28% 29% 29% 28% 28% 28% 29% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 56% 55% 62% 56% 56% 61% 56% 55% 63% Specificity 54% 54% 53% 53% 54% 52% 53% 53% 53% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 32% 32% 35% 33% 32% 37% 31% 30% 38% Specificity 80% 80% 77% 79% 79% 78% 78% 77% 78% OR Quartile 2 1.62 1.59 2.78 1.77 1.72 3.08 1.74 1.64 4.42 p Value 0.068 0.083 0.011 0.040 0.051 0.0081 0.056 0.084 0.0022 Lower limit of 95% CI 0.964 0.941 1.27 1.03 0.998 1.34 0.985 0.935 1.70 Upper limit of 95% CI 2.73 2.69 6.12 3.05 2.97 7.06 3.07 2.87 11.5 OR Quartile 3 1.48 1.43 1.78 1.47 1.49 1.75 1.46 1.36 1.93 p Value 0.082 0.12 0.042 0.096 0.086 0.053 0.11 0.20 0.025 Lower limit of 95% CI 0.951 0.916 1.02 0.935 0.945 0.993 0.919 0.854 1.09 Upper limit of 95% CI 2.29 2.22 3.11 2.30 2.33 3.08 2.32 2.15 3.45 OR Quartile 4 1.92 1.85 1.87 1.86 1.79 2.04 1.64 1.49 2.17 p Value 0.011 0.017 0.037 0.017 0.025 0.018 0.062 0.13 0.011 Lower limit of 95% CI 1.16 1.12 1.04 1.12 1.08 1.13 0.976 0.887 1.20 Upper limit of 95% CI 3.18 3.07 3.35 3.09 2.98 3.68 2.75 2.50 3.94

TABLE 16.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal staus is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatnine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3100 3510 3100 3520 3140 3620 Average 3400 4420 3460 4440 3550 4400 Stdev 1530 3080 1740 3060 2060 2870 p (t-test) 1.2E−4 2.8E−4 0.0021 Min 1080 863 1080 863 1080 863 Max 9100 19700 14500 19700 19700 18800 n (Patient) 176 151 190 137 205 122 sCr only Median 3100 3500 3100 3510 3130 3510 Average 3420 4420 3490 4410 3590 4330 Stdev 1590 3100 1780 3070 2100 2860 p (t-test) 2.2E−4 6.9E−4 0.0078 Min 1080 863 1080 863 1080 863 Max 9820 19700 14500 19700 19700 18800 n (Patient) 182 144 194 132 207 119 UO only Median 3100 3960 3110 3960 3110 3990 Average 3510 5010 3540 5000 3530 5050 Stdev 2000 3190 2030 3200 2030 3200 p (t-test) 1.6E−6 3.9E−6 1.9E−6 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 249 76 253 72 254 71 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.58 0.65 0.59 0.58 0.65 0.58 0.57 0.65 SE 0.032 0.032 0.038 0.032 0.032 0.039 0.033 0.033 0.039 p Value 0.0079 0.012 8.4E−5 0.0071 0.013 1.6E−4 0.014 0.042 6.7E−5 nCohort Non-persistent 176 182 249 190 194 253 205 207 254 nCohort Persistent 151 144 76 137 132 72 122 119 71 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 80% 80% 88% 82% 81% 89% 82% 81% 90% Specificity 30% 29% 29% 30% 29% 29% 29% 29% 29% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 54% 53% 61% 54% 54% 60% 55% 54% 61% Specificity 53% 53% 53% 53% 53% 53% 53% 52% 53% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 31% 31% 36% 32% 32% 36% 31% 30% 37% Specificity 80% 80% 78% 80% 79% 78% 79% 78% 78% OR Quartile 2 1.69 1.63 3.03 1.92 1.78 3.24 1.88 1.66 3.76 p Value 0.045 0.065 0.0037 0.016 0.034 0.0032 0.025 0.068 0.0017 Lower limit of 95% CI 1.01 0.971 1.43 1.13 1.04 1.48 1.08 0.964 1.65 Upper limit of 95% CI 2.83 2.73 6.40 3.27 3.04 7.10 3.26 2.87 8.58 OR Quartile 3 1.30 1.28 1.73 1.31 1.29 1.64 1.36 1.27 1.71 p Value 0.24 0.27 0.040 0.24 0.26 0.067 0.18 0.30 0.049 Lower limit of 95% CI 0.839 0.828 1.03 0.840 0.828 0.966 0.865 0.808 1.00 Upper limit of 95% CI 2.00 1.99 2.92 2.03 2.01 2.80 2.13 1.99 2.93 OR Quartile 4 1.82 1.78 1.94 1.89 1.80 1.99 1.66 1.52 2.04 p Value 0.020 0.025 0.019 0.013 0.023 0.017 0.052 0.11 0.013 Lower limit of 95% CI 1.10 1.08 1.11 1.14 1.08 1.13 0.996 0.911 1.16 Upper limit of 95% CI 3.02 2.95 3.39 3.14 2.98 3.50 2.75 2.53 3.60

Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3090 3510 3100 3520 3120 3520 Average 3390 4400 3460 4420 3490 4470 Stdev 1540 3060 1740 3040 1740 3140 p (t-test) 1.5E−4 3.9E−4 3.6E−4 Min 1080 863 1080 863 1080 863 Max 9100 19700 14500 19700 14500 19700 n (Patient) 173 154 188 139 200 127 sCr only Median 3090 3510 3080 3620 3100 3620 Average 3390 4430 3450 4430 3480 4480 Stdev 1520 3110 1740 3060 1730 3170 p (t-test) 9.8E−5 2.9E−4 2.7E−4 Min 1080 863 1080 863 1080 863 Max 9100 19700 14500 19700 14500 19700 n (Patient) 179 147 190 136 202 124 UO only Median 3080 3990 3100 3960 3100 3990 Average 3480 4930 3510 4930 3510 4970 Stdev 2000 3080 2020 3110 2020 3110 p (t-test) 1.4E−6 4.0E−6 2.0E−6 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 240 85 245 80 246 79 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.58 0.65 0.58 0.59 0.64 0.58 0.58 0.65 SE 0.032 0.032 0.036 0.032 0.032 0.037 0.033 0.033 0.0037 p Value 0.0072 0.0081 3.0E−5 0.0087 0.0072 9.2E−5 0.017 0.015 3.8E−5 nCohort Non-persistent 173 179 240 188 190 245 200 202 246 nCohort Persistent 154 147 85 139 136 80 127 124 79 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 80% 80% 88% 81% 81% 89% 81% 81% 90% Specificity 29% 29% 30% 30% 29% 29% 29% 29% 30% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 54% 54% 61% 54% 54% 60% 54% 55% 61% Specificity 53% 53% 54% 53% 53% 53% 52% 53% 53% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 31% 31% 35% 32% 32% 36% 31% 31% 37% Specificity 80% 80% 78% 80% 79% 78% 78% 78% 78% OR Quartile 2 1.66 1.60 3.15 1.84 1.77 3.28 1.75 1.68 3.74 p Value 0.053 0.075 0.0017 0.023 0.035 0.0018 0.041 0.060 9.1E−4 Lower limit of 95% CI 0.994 0.954 1.54 1.09 1.04 1.56 1.02 0.978 1.72 Upper limit of 95% CI 2.77 2.67 6.45 3.13 3.00 6.92 3.01 2.88 8.17 OR Quartile 3 1.33 1.31 1.83 1.30 1.35 1.70 1.31 1.37 1.76 p Value 0.20 0.22 0.019 0.24 0.18 0.044 0.23 0.17 0.031 Lower limit of 95% CI 0.859 0.848 1.11 0.840 0.871 1.02 0.842 0.873 1.05 Upper limit of 95% CI 2.05 2.03 3.03 2.02 2.11 2.83 2.05 2.14 2.96 OR Quartile 4 1.73 1.81 1.97 1.83 1.79 2.06 1.62 1.59 2.11 p Value 0.033 0.021 0.014 0.019 0.024 0.0097 0.062 0.075 0.0076 Lower limit of 95% CI 1.04 1.09 1.15 1.10 1.08 1.19 0.976 0.955 1.22 Upper limit of 95% CI 2.87 3.00 3.39 3.03 2.97 3.56 2.68 2.64 3.66

TABLE 16.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3040 3520 3040 3770 3080 3720 Average 3370 4400 3430 4430 3480 4460 Stdev 1540 3020 1750 2990 1750 3100 p (t-test) 1.1E−4 2.1E−4 3.0E−4 Min 1080  863 1080  863 1080  863 Max 9100 19700  14500  19700  14500  19700  n (Patient)  169  158  183  144  196  131 sCr only Median 3040 3520 3040 3770 3050 3770 Average 3360 4450 3420 4450 3450 4500 Stdev 1510 3080 1740 3030 1730 3120 p (t-test) 4.2E−5 1.4E−4 1.3E−4 Min 1080  863 1080  863 1080  863 Max 9100 19700  14500  19700  14500  19700  n (Patient)  175  151  186  140  198  128 UO only Median 3040 4030 3040 4180 3040 4300 Average 3460 4920 3450 4970 3440 5000 Stdev 1990 3030 1990 3050 1980 3040 p (t-test) 6.1E−7 2.9E−7 1.4E−7 Min  893  863  893  863  893  863 Max 19700  18800  19700  18800  19700  18800  n (Patient)  235   90  237   88  238   87 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.59 0.60 0.66 0.60 0.60 0.66 0.59 0.59 0.67 SE  0.031  0.032  0.035  0.032  0.032  0.035  0.032  0.033  0.035 p Value   0.0032   0.0022 7.1E−6   0.0024   0.0019 3.7E−6   0.0084   0.0044 1.3E−6 nCohort 169      175      235      183      186      237      196      198      238      Non-persistent nCohort Persistent 158      151      90     144      140      88     131      128      87     Cutoff Quartile 2 2320       2320       2330       2320       2320       2330       2320       2320       2330       Sensitivity 80% 80% 89% 82% 81% 90% 82% 81% 91% Specificity 30% 30% 30% 31% 30% 30% 30% 29% 31% Cutoff Quartile 3 3280       3280       3280       3280       3280       3280       3280       3280       3280       Sensitivity 54% 55% 62% 55% 56% 62% 55% 56% 63% Specificity 54% 54% 54% 54% 54% 54% 53% 54% 55% Cutoff Quartile 4 4720       4710       4720       4720       4710       4720       4720       4710       4720       Sensitivity 30% 32% 36% 31% 32% 36% 30% 31% 37% Specificity 80% 81% 79% 80% 80% 79% 78% 79% 79% OR Quartile 2 1.77 1.71 3.46 2.00 1.89 3.83 1.87 1.80 4.37 p Value  0.029  0.042 6.5E−4  0.010  0.018 4.0E−4  0.022  0.033 2.0E−4 Lower limit 1.06 1.02 1.70 1.18 1.11 1.82 1.09 1.05 2.01 of 95% CI Upper limit 2.95 2.85 7.07 3.39 3.20 8.05 3.21 3.08 9.51 of 95% CI OR Quartile 3 1.39 1.45 1.97 1.40 1.49 1.99 1.38 1.51 2.07 p Value 0.14  0.096   0.0075 0.13  0.074   0.0071 0.16  0.070   0.0048 Lower limit  0.902  0.936 1.20  0.904  0.962 1.21  0.885  0.967 1.25 of 95% CI Upper limit 2.15 2.24 3.24 2.17 2.32 3.29 2.15 2.36 3.43 of 95% CI OR Quartile 4 1.73 1.93 2.04 1.79 1.91 2.14 1.51 1.69 2.19 p Value  0.033  0.011   0.0087  0.023  0.012   0.0054 0.11  0.042   0.0042 Lower limit 1.04 1.16 1.20 1.08 1.15 1.25  0.911 1.02 1.28 of 95% CI Upper limit 2.87 3.21 3.48 2.97 3.16 3.65 2.50 2.80 3.74 of 95% CI

Example 17. Use of Metalloproteinase Inhibitor 4 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. Metalloproteinase inhibitor 4 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 17.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.7 36.2 15.6 37.0 15.4 42.2 Average 26.9 239   34.3 284   35.2 320   Stdev 45.9 969   57.0 1080    59.2 1150    p (t-test)    0.018     0.0039     0.0010 Min      0.00110      0.00167      0.00110      0.00167      0.00110      0.00167 Max 409   9030    409   9030    409   9030    n (Patient) 119   198   158   159   179   138   sCr only Median 13.3 35.4 16.0 37.0 16.0 42.2 Average 29.3 250   31.2 294   33.1 336   Stdev 46.8 999   46.8 1090    51.4 1180    p (t-test)    0.012     0.0024 5.6E−4 Min      0.00110      0.00167      0.00110      0.00167      0.00110      0.00167 Max 409   9030    409   9030    409   9030    n (Patient) 130   186   162   154   184   132   UO only Median 18.2 41.0 19.1 53.0 19.3 57.9 Average 96.3 368   99.9 422   99.5 467   Stdev 598   1160    588   1280    581   1360    p (t-test)     0.0081     0.0038     0.0017 Min      0.00110   1.76      0.00110   1.76      0.00110 1.76 Max 9030    7570    9030    7570    9030    7570    n (Patient) 241   74  256   59  263   52  Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.69 0.67 0.68 0.67 0.67 0.69 0.69 0.69 0.70 SE  0.029  0.030  0.038  0.030  0.030  0.041  0.030  0.031  0.043 p Value 9.3E−11 2.2E−8 2.6E−6 2.2E−8 2.5E−8 3.9E−6 1.1E−10 2.2E−10 2.0E−6 nCohort 119      130      241     158      162      256      179      184      263      Non- persistent nCohort 198      186      74     159      154      59   138      132      52     Persistent Cutoff 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Quartile 2 Sensitivity 82% 81% 88% 82% 81% 88% 86% 85% 88% Specificity 36% 33% 29% 32% 31% 28% 33% 32% 28% Cutoff 24.1   23.6   24.1   24.1   23.6   24.1   24.1   23.6   24.1   Quartile 3 Sensitivity 60% 59% 69% 60% 60% 73% 64% 64% 77% Specificity 66% 63% 56% 60% 60% 55% 61% 60% 55% Cutoff 60.9   60.8   61.8   60.9   60.8   61.8   60.9   60.8   61.8   Quartile 4 Sensitivity 35% 35% 42% 38% 40% 44% 41% 42% 46% Specificity 92% 90% 80% 88% 89% 79% 87% 87% 79% OR 2.55 2.06 2.96 2.23 1.92 2.91 2.90 2.64 2.95 Quartile 2 p Value 4.3E−4   0.0060   0.0046   0.0029  0.014  0.012 2.3E−4 7.9E−4  0.018 Lower 1.51 1.23 1.40 1.32 1.14 1.26 1.65 1.50 1.21 limit of 9 5% CI Upper 4.28 3.45 6.26 3.78 3.25 6.70 5.12 4.66 7.19 limit of 95% CI OR 2.81 2.47 2.78 2.30 2.28 3.30 2.83 2.75 4.10 Quartile 3 p Value 1.9E−5 1.2E−4 3.0E−4 2.9E−4 3.5E−4 1.8E−4 9.5E−6 1.8E−5 6.1E−5 Lower 1.75 1.56 1.60 1.47 1.45 1.76 1.79 1.73 2.06 limit of 9 5% CI Upper 4.50 3.92 4.83 3.60 3.57 6.15 4.48 4.37 8.16 limit of 95% CI OR 5.96 4.95 2.90 4.55 5.25 3.02 4.41 4.76 3.24 Quartile 4 p Value 8.3E−7 1.3E−6 1.9E−4 2.5E−7 3.1E−8 2.8E−4 1.1E−7 2.9E−8 2.1E−4 Lower 2.93 2.59 1.66 2.56 2.92 1.66 2.55 2.74 1.74 limit of 95% CI Upper 12.1   9.45 5.07 8.10 9.44 5.48 7.63 8.26 6.03 limit of 95% CI

TABLE 17.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.5 34.8 14.7 37.0 13.3 41.1 Average 27.2 231    30.5 271    31.7 298    Stdev 47.1 953    48.1 1040     52.4 1100     p (t-test)    0.024     0.0056     0.0021 Min      0.00110      0.00167      0.00110      0.00167      0.00110      0.00167 Max 409    9030     409    9030     409    9030     n (Patient) 112    205    147    170    165    152    sCr only Median 13.3 34.9 16.3 36.9 15.4 40.1 Average 29.5 246    31.6 281    32.9 305    Stdev 47.4 989    47.8 1070     52.3 1120     p (t-test)    0.015     0.0041     0.0017 Min      0.00110      0.00167      0.00110      0.00167      0.00110      0.00167 Max 409    9030     409    9030     409    9030     n (Patient) 126    190    154    162    169    147    UO only Median 18.1 39.3 18.2 40.3 18.2 47.4 Average 99.3 306    103    338    102    363    Stdev 622    1040     609    1130     601    1180     p (t-test)    0.031    0.020    0.013 Min      0.00110      0.00277      0.00110      0.00277      0.00110      0.00277 Max 9030     7570     9030     7570     9030     7570     n (Patient) 222    93   238    77   245    70   Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.69 0.66 0.66 0.68 0.66 0.66 0.70 0.68 0.67 SE  0.030  0.030  0.035  0.030  0.030  0.037  0.029  0.030  0.039 p Value 2.2E−10 5.2E−8 4.9E−6 1.6E−9 9.6E−8 1.4E−5 3.0E−12 1.4E−9 1.1E−5 nCohort 112      126      222      147      154      238      165      169      245      Non- persistent nCohort 205      190      93     170      162      77     152      147      70     Persistent Cutoff 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Quartile 2 Sensitivity 82% 81% 87% 83% 81% 87% 86% 84% 87% Specificity 38% 33% 30% 34% 32% 29% 35% 33% 29% Cutoff 24.1   23.6   24.1   24.1   23.6   24.1   24.1   23.6   24.1   Quartile 3 Sensitivity 59% 58% 67% 61% 59% 69% 64% 62% 71% Specificity 66% 63% 57% 62% 60% 56% 62% 60% 56% Cutoff 60.9   60.8   61.8   60.9   60.8   61.8   60.9   60.8   61.8   Quartile 4 Sensitivity 34% 35% 38% 38% 38% 39% 39% 39% 40% Specificity 91% 90% 80% 89% 88% 79% 88% 87% 79% OR 2.72 2.07 2.92 2.51 2.05 2.74 3.38 2.67 2.71 Quartile 2 p Value 1.7E−4   0.0058   0.0017 6.1E−4   0.0069   0.0062 2.1E−5 4.4E−4   0.0094 Lower 1.62 1.23 1.49 1.48 1.22 1.33 1.93 1.54 1.28 limit of 9 5% CI Upper 4.59 3.46 5.71 4.24 3.46 5.63 5.92 4.62 5.76 limit of 95% CI OR 2.81 2.36 2.63 2.50 2.16 2.80 2.93 2.47 3.17 Quartile 3 p Value 2.5E−5 2.7E−4 1.9E−4 7.5E−5 7.9E−4 2.2E−4 4.0E−6 9.1E−5 8.8E−5 Lower 1.74 1.49 1.58 1.59 1.38 1.62 1.86 1.57 1.78 limit of 95% CI Upper 4.53 3.75 4.36 3.93 3.38 4.83 4.63 3.89 5.65 limit of 95% CI OR 5.29 4.63 2.44 4.94 4.56 2.46 4.73 4.23 2.54 Quartile 4 p Value 4.4E−6 3.5E−6   0.0010 2.2E−7 3.7E−7   0.0015 9.0E−8 3.9E−7   0.0013 Lower 2.60 2.42 1.43 2.70 2.54 1.41 2.68 2.42 1.44 limit of 95% CI Upper 10.8   8.84 4.16 9.05 8.19 4.29 8.36 7.39 4.48 limit of 95% CI

TABLE 17.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.3 34.7 13.3 36.7 13.3 37.0 Average 27.2 230   30.6 266   32.4 285   Stdev 47.3 951   48.6 1030    53.4 1080    p (t-test)    0.025     0.0066     0.0034 Min      0.00110      0.00167      0.00110      0.00167      0.00110      0.00167 Max 409   9030    409   9030    409   9030    n (Patient) 111   206   144   173   158   159   sCr only Median 13.3 34.7 16.3 36.5 15.4 37.0 Average 29.7 243   31.8 276   33.6 293   Stdev 47.7 984   48.2 1060    53.1 1100    p (t-test)    0.016     0.0049     0.0027 Min      0.00110      0.00167      0.00110      0.00167      0.00110      0.00167 Max 409   9030    409   9030    409   9030    n (Patient) 124   192   151   165   163   153   UO only Median 18.1 37.0 18.2 39.1 18.5 39.3 Average 102   279   106   295   106   312   Stdev 637   995   626   1050    617   1090    p (t-test)    0.057    0.051    0.037 Min      0.00110      0.00277      0.00110      0.00277      0.00110      0.00277 Max 9030    7570    9030    7570    9030    7570    n (Patient) 212   103   225   90  232   83  Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.66 0.64 0.68 0.66 0.64 0.69 0.67 0.64 SE  0.030  0.030  0.034  0.030  0.030  0.036  0.030  0.030  0.037 p Value 1.8E−10 8.1E−8 5.9E−5 1.0E−9 1.4E−7 1.5E−4 1.2E−10 2.2E−8 1.6E−4 nCohort 111      124      212      144      151      225      158      163      232      Non- persistent nCohort 206      192      103      173      165      90     159      153      83     Persistent Cutoff 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Quartile 2 Sensitivity 82% 81% 85% 83% 82% 86% 85% 84% 86% Specificity 38% 34% 30% 35% 32% 29% 35% 33% 29% Cutoff 24.1   23.6   24.1   24.1   23.6   24.1   24.1   23.6   24.1   Quartile 3 Sensitivity 59% 58% 63% 61% 59% 64% 63% 61% 66% Specificity 67% 63% 56% 63% 60% 56% 63% 60% 56% Cutoff 60.9   60.8   61.8   60.9   60.8   61.8   60.9   60.8   61.8   Quartile 4 Sensitivity 34% 34% 35% 37% 37% 36% 38% 37% 36% Specificity 91% 90% 80% 89% 88% 79% 87% 87% 79% OR 2.78 2.15 2.54 2.64 2.16 2.46 3.00 2.54 2.40 Quartile 2 p Value 1.3E−4   0.0038   0.0033 3.0E−4   0.0038   0.0070 7.4E−5 7.1E−4  0.011 Lower 1.65 1.28 1.36 1.56 1.28 1.28 1.74 1.48 1.22 limit of 95% CI Upper 4.69 3.60 4.72 4.47 3.64 4.73 5.17 4.35 4.72 limit of 95% CI OR 2.90 2.37 2.19 2.72 2.22 2.27 2.84 2.34 2.46 Quartile 3 p Value 1.5E−5 2.6E−4   0.0015 1.8E−5 5.2E−4   0.0015 6.9E−6 2.3E−4 7.5E−4 Lower 1.79 1.49 1.35 1.72 1.41 1.37 1.80 1.49 1.46 limit of 95% CI Upper 4.71 3.77 3.55 4.29 3.48 3.76 4.48 3.67 4.15 limit of 95% CI OR 5.20 4.47 2.11 4.70 4.33 2.09 4.18 3.81 2.11 Quartile 4 p Value 5.5E−6 5.7E−6   0.0053 5.3E−7 9.0E−7   0.0073 8.0E−7 2.5E−6   0.0074 Lower 2.55 2.34 1.25 2.57 2.41 1.22 2.37 2.18 1.22 limit of 95% CI Upper 10.6   8.54 3.57 8.60 7.78 3.58 7.38 6.64 3.66 limit of 95% CI

TABLE 17.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.5 34.6 13.3 36.6 13.3 37.0 Average 27.4 229 30.8 265 32.5 282 Stdev 47.5 949 48.7 1030 53.7 1070 p (t-test) 0.027 0.0071 0.0039 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 110 207 143 174 156 161 sCr only Median 13.3 34.7 16.3 34.9 15.9 36.9 Average 29.7 243 32.0 275 33.7 292 Stdev 47.7 984 48.4 1060 53.3 1090 p (t-test) 0.016 0.0052 0.0029 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 124 192 150 166 162 154 UO only Median 18.2 36.7 18.2 37.0 18.2 38.8 Average 104 270 108 281 108 294 Stdev 643 977 633 1030 625 1060 p (t-test) 0.071 0.068 0.055 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 208 107 220 95 226 89 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.69 0.66 0.63 0.68 0.66 0.63 0.69 0.67 0.63 SE 0.030 0.030 0.034 0.030 0.030 0.035 0.030 0.030 0.036 p Value 6.1E−10 8.1E−8 1.5E−4 2.3E−9 2.7E−7 1.9E−4 1.9E−10 4.7E−8 1.7E−4 nCohort Non- 110 124 208 143 150 220 156 162 226 persistent nCohort Persistent 207 192 107 174 166 95 161 154 89 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 82% 81% 84% 83% 81% 85% 84% 83% 85% Specificity 37% 34% 30% 34% 32% 30% 35% 33% 29% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 59% 58% 62% 61% 59% 63% 63% 60% 65% Specificity 66% 63% 56% 63% 60% 55% 63% 60% 56% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 34% 34% 35% 37% 37% 35% 37% 37% 35% Specificity 91% 90% 80% 89% 88% 79% 87% 86% 79% OR Quartile 2 2.64 2.15 2.25 2.50 2.05 2.43 2.88 2.39 2.41 p Value 2.7E−4 0.0038 0.0079 5.9E−4 0.0068 0.0064 1.2E−4 0.0014 0.0084 Lower limit of 1.57 1.28 1.24 1.48 1.22 1.28 1.68 1.40 1.25 95% CI Upper limit of 4.46 3.60 4.09 4.22 3.45 4.59 4.94 4.08 4.64 95% CI OR Quartile 3 2.83 2.37 2.03 2.65 2.16 2.13 2.84 2.28 2.36 p Value 2.4E−5 2.6E−4 0.0036 2.9E−5 7.8E−4 0.0027 6.9E−6 3.5E−4 9.6E−4 Lower limit of 1.75 1.49 1.26 1.68 1.38 1.30 1.80 1.45 1.42 95% CI Upper limit of 4.59 3.77 3.27 4.18 3.39 3.50 4.49 3.57 3.92 95% CI OR Quartile 4 5.11 4.47 2.09 4.62 4.26 2.01 4.04 3.74 1.98 p Value 6.9E−6 5.7E−6 0.0057 7.0E−7 1.2E−6 0.010 1.4E−6 3.3E−6 0.013 Lower limit of 2.51 2.34 1.24 2.52 2.37 1.18 2.29 2.14 1.15 95% CI Upper limit of 10.4 8.54 3.52 8.45 7.65 3.43 7.13 6.52 3.40 95% CI

TABLE 17.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (OU) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.5 34.6 14.0 35.9 13.3 36.7 Average 26.9 226 30.6 259 32.6 273 Stdev 47.5 940 49.0 1020 54.2 1050 p (t-test) 0.030 0.0090 0.0055 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 106 211 138 179 150 167 sCr only Median 13.3 34.8 15.9 36.2 15.1 36.9 Average 29.1 240 31.5 269 33.3 286 Stdev 47.7 977 48.5 1040 53.5 1080 p (t-test) 0.018 0.0063 0.0036 Min 0.00110 0.00167 0.00110 0.00167 0.00110 0.00167 Max 409 9030 409 9030 409 9030 n (Patient) 121 195 146 170 158 158 UO only Median 17.7 37.0 18.1 37.0 18.2 37.9 Average 97.1 273 103 279 103 287 Stdev 648 955 641 990 634 1010 p (t-test) 0.053 0.058 0.050 Min 0.00110 0.00277 0.00110 0.00277 0.00110 0.00277 Max 9030 7570 9030 7570 9030 7570 n (Patient) 202 113 212 103 217 98 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.69 0.67 0.65 0.67 0.66 0.64 0.68 0.67 0.64 SE 0.030 0.030 0.033 0.030 0.030 0.034 0.030 0.030 0.035 p Value 5.2E−10 2.6E−8 1.1E−5 5.1E−9 8.2E−8 2.9E−5 7.4E−10 1.4E−8 3.8E−5 nCohort Non- 106 121 202 138 146 212 150 158 217 persistent nCohort Persistent 211 195 113 179 170 103 167 158 98 Cutoff Quartile 2 8.33 8.31 8.36 8.33 8.31 8.36 8.33 8.31 8.36 Sensitivity 82% 81% 85% 83% 82% 86% 84% 84% 87% Specificity 38% 35% 31% 35% 33% 31% 35% 34% 30% Cutoff Quartile 3 24.1 23.6 24.1 24.1 23.6 24.1 24.1 23.6 24.1 Sensitivity 59% 58% 63% 60% 59% 63% 62% 61% 64% Specificity 67% 64% 57% 63% 61% 56% 63% 61% 56% Cutoff Quartile 4 60.9 60.8 61.8 60.9 60.8 61.8 60.9 60.8 61.8 Sensitivity 34% 34% 35% 36% 36% 35% 37% 37% 35% Specificity 92% 90% 81% 89% 88% 80% 87% 87% 79% OR Quartile 2 2.67 2.27 2.50 2.55 2.20 2.81 2.96 2.56 2.86 p Value 2.4E−4 0.0019 0.0026 4.5E−4 0.0030 0.0014 7.1E−5 5.6E−4 0.0016 Lower limit of 1.58 1.35 1.38 1.51 1.31 1.49 1.73 1.50 1.49 95% CI Upper limit of 4.52 3.81 4.54 4.29 3.70 5.30 5.06 4.37 5.48 95% CI OR Quartile 3 2.89 2.46 2.23 2.59 2.29 2.19 2.70 2.40 2.31 p Value 2.1E−5 1.6E−4 8.5E−4 4.4E−5 3.4E−4 0.0015 1.8E−5 1.5E−4 8.6E−4 Lower limit of 1.77 1.54 1.39 1.64 1.45 1.35 1.71 1.53 1.41 95% CI Upper limit of 4.71 3.93 3.58 4.10 3.59 3.55 4.26 3.77 3.78 95% CI OR Quartile 4 5.47 4.75 2.29 4.68 4.36 2.11 3.97 3.78 2.03 p Value 6.9E−6 4.4E−6 0.0018 9.4E−7 1.2E−6 0.0053 2.6E−6 3.4E−6 0.0088 Lower limit of 2.61 2.44 1.36 2.52 2.40 1.25 2.23 2.16 1.20 95% CI Upper limit of 11.5 9.25 3.85 8.66 7.89 3.57 7.05 6.64 3.45 95% CI

TABLE 17.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2870 3560 3000 3640 3110 3520 Average 3210 4220 3360 4330 3520 4290 Stdev 1650 2690 1670 2880 1860 2920 p (t-test) 2.8E−4 2.5E−4 0.0044 Min 1080 863 1080 863 1080 863 Max 9820 19700 9820 19700 14500 19700 n (Patient) 114 213 155 172 179 148 sCr only Median 2930 3510 2950 3560 3070 3670 Average 3290 4210 3310 4370 3460 4360 Stdev 1640 2750 1580 2920 1810 2960 p (t-test) 7.5E−4 6.7E−5 8.3E−4 Min 1080 863 1080 863 1080 863 Max 9820 19700 9820 19700 14500 19700 n (Patient) 124 202 157 169 182 144 UO only Median 3040 4350 3100 4340 3100 4390 Average 3520 4910 3570 5040 3570 5240 Stdev 2050 3060 2050 3280 2050 3390 p (t-test) 6.0E−6 1.0E−5 1.8E−6 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 245 80 261 64 269 56 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.63 0.61 0.65 0.61 0.61 0.64 0.57 0.59 0.66 SE 0.031 0.031 0.037 0.031 0.031 0.040 0.032 0.032 0.042 p Value 3.1E−5 6.1E−4 2.8E−5 5.8E−4 2.9E−4 3.6E−4 0.027 0.0057 1.3E−4 nCohort Non- 114 124 245 155 157 261 179 182 269 persistent nCohort Persistent 213 202 80 172 169 64 148 144 56 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 81% 81% 85% 81% 82% 86% 80% 80% 88% Specificity 37% 35% 28% 32% 32% 28% 29% 29% 28% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 56% 55% 66% 56% 56% 64% 55% 56% 66% Specificity 61% 58% 55% 57% 57% 53% 54% 54% 53% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 31% 31% 39% 32% 33% 36% 30% 33% 38% Specificity 86% 85% 79% 83% 83% 77% 79% 81% 77% OR Quartile 2 2.52 2.22 2.22 2.08 2.14 2.33 1.61 1.63 2.66 p Value 4.1E−4 0.0021 0.020 0.0049 0.0036 0.028 0.069 0.065 0.022 Lower limit of 1.51 1.33 1.13 1.25 1.28 1.09 0.963 0.971 1.15 95% CI Upper limit of 4.21 3.69 4.36 3.47 3.58 4.95 2.69 2.73 6.13 95% CI OR Quartile 3 2.05 1.69 2.41 1.70 1.68 2.03 1.40 1.49 2.21 p Value 0.0024 0.023 0.0011 0.018 0.020 0.014 0.13 0.075 0.0099 Lower limit of 1.29 1.08 1.42 1.10 1.08 1.15 0.903 0.961 1.21 95% CI Upper limit of 3.27 2.65 4.08 2.63 2.60 3.58 2.16 2.31 4.04 95% CI OR Quartile 4 2.75 2.50 2.41 2.23 2.50 1.92 1.68 2.04 2.05 p Value 0.0010 0.0017 0.0016 0.0027 7.0E−4 0.029 0.044 0.0060 0.022 Lower limit of 1.50 1.41 1.39 1.32 1.47 1.07 1.01 1.23 1.11 95% CI Upper limit of 5.03 4.44 4.15 3.76 4.24 3.46 2.77 3.38 3.77 95% CI

TABLE 17.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2820 3560 2910 3720 3080 3520 Average 3120 4240 3230 4370 3450 4300 Stdev 1530 2690 1500 2860 1780 2880 p (t-test) 6.7E−5 1.8E−5 0.0015 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 108 219 144 183 164 163 sCr only Median 2900 3510 2950 3520 3100 3500 Average 3260 4210 3290 4340 3490 4240 Stdev 1640 2730 1590 2870 1860 2860 p (t-test) 5.8E−4 9.0E−5 0.0052 Min 1080 863 1080 863 1080 863 Max 9820 19700 9820 19700 14500 19700 n (Patient) 120 206 149 177 166 160 UO only Median 3030 3960 3070 3930 3080 3930 Average 3460 4760 3520 4830 3530 4930 Stdev 2020 2930 2030 3090 2020 3150 p (t-test) 5.6E−6 1.5E−5 5.9E−6 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 1880 n (Patient) 225 100 241 84 248 77 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.65 0.61 0.65 0.63 0.61 0.63 0.58 0.57 0.64 SE 0.031 0.031 0.034 0.031 0.031 0.037 0.031 0.032 0.038 p Value 2.9E−6 2.6E−4 1.6E−5 2.8E−5 2.2E−4 2.6E−4 0.011 0.022 1.5E−4 nCohort Non- 108 120 225 144 149 241 164 166 248 persistent nCohort Persistent 219 206 100 183 177 84 163 160 77 Cutoff Quartile 2 2320 2320 2330 2320 2320 2330 2320 2320 2330 Sensitivity 82% 81% 86% 82% 82% 87% 80% 80% 88% Specificity 39% 36% 31% 35% 34% 29% 30% 30% 29% Cutoff Quartile 3 3280 3280 3280 3280 3280 3280 3280 3280 3280 Sensitivity 57% 55% 64% 57% 56% 62% 55% 54% 62% Specificity 63% 59% 56% 58% 57% 54% 54% 54% 54% Cutoff Quartile 4 4720 4710 4720 4720 4710 4720 4720 4710 4720 Sensitivity 31% 31% 36% 33% 32% 33% 31% 30% 34% Specificity 87% 85% 80% 85% 83% 78% 80% 80% 77% OR Quartile 2 2.85 2.39 2.60 2.34 2.29 2.72 1.68 1.72 3.09 p Value 7.2E−5 8.2E−4 0.0030 0.0011 0.0015 0.0047 0.046 0.036 0.0031 Lower limit of 1.70 1.44 1.38 1.41 1.37 1.36 1.01 1.04 1.46 95% CI Upper limit of 4.78 3.98 4.91 3.91 3.82 5.43 2.79 2.87 6.53 95% CI OR Quartile 3 2.22 1.80 2.26 1.84 1.69 1.90 1.43 1.34 1.91 p Value 9.6E−4 0.012 9.9E−4 0.0067 0.020 0.013 0.11 0.18 0.015 Lower limit of 1.38 1.14 1.39 1.18 1.09 1.15 0.924 0.869 1.13 95% CI Upper limit of 3.56 2.83 3.68 2.87 2.62 3.16 2.21 2.08 3.23 95% CI OR Quartile 4 3.02 2.55 2.19 2.71 2.36 1.73 1.83 1.66 1.75 p Value 5.8E−4 0.0016 0.0032 3.8E−4 0.0016 0.049 0.021 0.049 0.050 Lower limit of 1.61 1.43 1.30 1.56 1.38 1.00 1.10 1.00 1.00 95% CI Upper limit of 5.68 4.57 3.69 4.68 4.01 2.99 3.04 2.76 3.05 95% CI

TABLE 17.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2800 3540 2910 3560 3100 3490 Average 3120 4240 3230 4360 3440 4270 Stdev 1540 2680 1510 2850 1800 2840 p (t-test) 7.6E−5 2.7E−5 0.0021 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 107 220 142 185 158 169 sCr only Median 2870 3510 2930 3520 3100 3480 Average 3210 4230 3260 4350 3440 4270 Stdev 1540 2740 1510 2880 1790 2860 p (t-test) 2.1E−4 4.0E−5 0.0021 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 118 208 146 180 160 166 UO only Median 3010 3920 3100 3720 3100 3790 Average 3440 4700 3520 4690 3530 4770 Stdev 2020 2870 2020 3020 2020 3070 p (t-test) 6.4E−6 6.1E−5 3.1E−5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 216 109 230 95 237 88 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.62 0.65 0.63 0.62 0.62 0.58 0.58 0.62 SE 0.031 0.031 0.033 0.031 0.031 0.035 0.031 0.032 0.036 p Value 3.1E−6 1.1E−4 1.1E−5 4.0E−5 1.3E−4 8.9E−4 0.010 0.012 6.2E−4 nCohort Non- 107 118 216 142 146 230 158 160 237 persistent nCohort 220 208 109 185 180 95 169 166 88 Persistent Cutoff Quartile 2320 2320 2330 2320 2320 2330 2320 2320 2330 2 Sensitivity 82% 81% 86% 82% 82% 85% 80% 81% 86% Specificity 39% 36% 31% 35% 34% 29% 31% 31% 29% Cutoff Quartile 3280 3280 3280 3280 3280 3280 3280 3280 3280 3 Sensitivity 56% 55% 61% 56% 56% 58% 53% 53% 58% Specificity 63% 59% 56% 58% 57% 53% 53% 53% 53% Cutoff Quartile 4720 4710 4720 4720 4710 4720 4720 4710 4720 4 Sensitivity 31% 31% 36% 32% 32% 33% 30% 30% 33% Specificity 87% 86% 80% 85% 84% 78% 80% 80% 78% OR Quartile 2 2.91 2.48 2.76 2.43 2.41 2.38 1.85 1.90 2.60 p Value 5.3E−5 4.9E−4 0.0013 6.8E−4 7.8E−4 0.0075 0.017 0.013 0.0052 Lower limit of 1.73 1.49 1.49 1.46 1.44 1.26 1.11 1.14 1.33 95% CI Upper limit of 4.88 4.14 5.11 4.05 4.02 4.49 3.08 3.17 5.08 95% CI OR Quartile 3 2.16 1.80 1.99 1.75 1.65 1.55 1.29 1.28 1.54 p Value 0.0014 0.012 0.0040 0.013 0.026 0.074 0.25 0.27 0.088 Lower limit of 1.35 1.14 1.25 1.13 1.06 0.959 0.837 0.828 0.939 95% CI Upper limit of 3.47 2.85 3.19 2.73 2.56 2.52 2.00 1.98 2.52 95% CI OR Quartile 4 2.97 2.70 2.24 2.62 2.42 1.70 1.77 1.72 1.71 p Value 7.1E−4 1.0E−3 0.0021 5.9E−4 0.0013 0.050 0.029 0.036 0.052 Lower limit of 1.58 1.49 1.34 1.51 1.41 1.00 1.06 1.04 0.995 95% CI Upper limit of 5.58 4.88 3.75 4.53 4.14 2.89 2.95 2.87 2.93 95% CI

TABLE 17.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2820 3520 2910 3560 3090 3500 Average 3130 4230 3240 4340 3440 4250 Stdev 1540 2680 1520 2840 1810 2820 p (t-test) 1.1E−4 3.6E−5 0.0024 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 106 221 140 187 155 172 sCr only Median 2870 3510 2930 3520 3100 3480 Average 3210 4230 3260 4340 3450 4250 Stdev 1540 2740 1520 2870 1800 2850 p (t-test) 2.1E−4 5.4E−5 0.0027 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 118 208 144 182 158 168 UO only Median 3010 3880 3070 3790 3070 3890 Average 3450 4610 3490 4670 3490 4740 Stdev 2040 2840 2020 2960 2020 2990 p (t-test) 2.8E−5 3.5E−5 1.7E−5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 211 114 223 102 229 96 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.62 0.63 0.62 0.62 0.62 0.58 0.58 0.63 SE 0.031 0.031 0.033 0.031 0.031 0.034 0.031 0.032 0.035 p Value 6.5E−6 1.1E−4 5.7E−5 5.1E−5 1.7E−4 3.4E−4 0.0094 0.014 1.7E−4 nCohort Non- 106 118 211 140 144 223 155 158 229 persistent nCohort 221 208 114 187 182 102 172 168 96 Persistent Cutoff Quartile 2320 2320 2330 2320 2320 2330 2320 2320 2330 2 Sensitivity 81% 81% 85% 82% 82% 85% 80% 80% 86% Specificity 39% 36% 30% 34% 34% 30% 31% 31% 30% Cutoff Quartile 3280 3280 3280 3280 3280 3280 3280 3280 3280 3 Sensitivity 56% 55% 60% 56% 55% 58% 53% 53% 58% Specificity 62% 59% 55% 58% 57% 53% 54% 53% 53% Cutoff Quartile 4720 4710 4720 4720 4710 4720 4720 4710 4720 4 Sensitivity 31% 31% 34% 32% 32% 33% 30% 30% 33% Specificity 87% 86% 80% 84% 83% 78% 80% 80% 78% OR Quartile 2 2.77 2.48 2.48 2.35 2.33 2.44 1.82 1.84 2.70 p Value 1.1E−4 4.9E−4 0.0026 0.0010 0.0012 0.0048 0.020 0.019 0.0028 Lower limit of 1.65 1.49 1.37 1.41 1.40 1.31 1.10 1.11 1.41 95% CI Upper limit of 4.65 4.14 4.50 3.91 3.88 4.53 3.02 3.06 5.16 95% CI OR Quartile 3 2.11 1.80 1.81 1.76 1.65 1.57 1.33 1.28 1.60 p Value 0.0020 0.012 0.012 0.012 0.026 0.062 0.20 0.27 0.057 Lower limit of 1.31 1.14 1.14 1.13 1.06 0.978 0.858 0.828 0.986 95% CI Upper limit of 3.39 2.85 2.87 2.74 2.56 2.52 2.05 1.98 2.58 95% CI OR Quartile 4 2.92 2.70 2.03 2.53 2.34 1.82 1.69 1.67 1.79 p Value 8.7E−4 1.0E−3 0.0066 9.0E−4 0.0020 0.024 0.045 0.049 0.031 Lower limit of 1.55 1.49 1.22 1.46 1.37 1.08 1.01 1.00 1.06 95% CI Upper limit of 5.49 4.88 3.39 4.39 4.00 3.07 2.81 2.78 3.03 95% CI

TABLE 17.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2850 3510 2920 3520 3090 3480 Average 3150 4200 3240 4310 3440 4230 Stdev 1570 2670 1540 2810 1830 2790 p (t-test) 2.7E−4 8.1E−5 0.0034 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 102 225 135 192 149 178 sCr only Median 2920 3500 2980 3510 3120 3450 Average 3230 4210 3270 4310 3460 4220 Stdev 1550 2730 1530 2850 1810 2830 p (t-test) 4.6E−4 1.1E−4 0.0048 Min 1080 863 1080 863 1080 863 Max 9100 19700 9100 19700 14500 19700 n (Patient) 115 211 140 186 154 172 UO only Median 3010 3910 3050 3860 3040 3890 Average 3430 4580 3470 4620 3470 4670 Stdev 2030 2810 2010 2900 2020 2910 p (t-test) 2.7E−5 3.4E−5 2.0E−5 Min 893 863 893 863 893 863 Max 19700 18800 19700 18800 19700 18800 n (Patient) 204 121 214 111 219 106 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.61 0.63 0.62 0.61 0.62 0.58 0.57 0.63 SE 0.032 0.032 0.032 0.031 0.031 0.033 0.031 0.032 0.034 p Value 2.8E−5 3.8E−4 4.3E−5 1.0E−4 3.8E−4 2.2E−4 0.010 0.024 1.0E−4 nCohort Non- 102 115 204 135 140 214 149 154 219 persistent nCohort 225 211 121 192 186 111 178 172 106 Persistent Cutoff 2320 2320 2330 2320 2320 2330 2320 2320 2330 Quartile 2 Sensitivity 81% 81% 84% 82% 81% 85% 80% 80% 86% Specificity 38% 36% 30% 35% 34% 30% 32% 31% 30% Cutoff 3280 3280 3280 3280 3280 3280 3280 3280 3280 Quartile 3 Sensitivity 55% 55% 60% 55% 55% 58% 53% 52% 58% Specificity 61% 58% 55% 57% 56% 54% 53% 53% 54% Cutoff 4720 4710 4720 4720 4710 4720 4720 4710 4720 Quartile 4 Sensitivity 30% 31% 34% 31% 31% 33% 29% 29% 33% Specificity 86% 85% 80% 84% 83% 79% 80% 79% 79% OR Quartile 2 2.62 2.30 2.34 2.40 2.18 2.36 1.88 1.72 2.62 p Value 2.8E−4 0.0014 0.0036 7.8E−4 0.0026 0.0046 0.014 0.036 0.0023 Lower limit of 1.56 1.38 1.32 1.44 1.31 1.30 1.14 1.04 1.41 95% CI Upper limit of 4.41 3.83 4.16 3.99 3.62 4.27 3.12 2.85 4.85 95% CI OR Quartile 3 1.90 1.67 1.82 1.64 1.57 1.58 1.26 1.22 1.65 p Value 0.0081 0.028 0.0097 0.030 0.044 0.052 0.29 0.37 0.037 Lower limit of 1.18 1.06 1.16 1.05 1.01 0.996 0.817 0.788 1.03 95% CI Upper limit of 3.06 2.65 2.88 2.55 2.45 2.51 1.95 1.88 2.63 95% CI OR Quartile 4 2.72 2.57 2.04 2.33 2.19 1.88 1.64 1.56 1.80 p Value 0.0019 0.0018 0.0061 0.0025 0.0043 0.016 0.060 0.086 0.025 Lower limit of 1.45 1.42 1.22 1.35 1.28 1.12 0.979 0.938 1.08 95% CI Upper limit of 5.12 4.64 3.39 4.04 3.75 3.14 2.74 2.60 3.03 95% CI

Example 18. Use of C—C Motif Chemokine 15 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 15 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 18.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.0 289 10.3 274 0.0534 237 Average 144 2060 118 1990 120 1950 Stdev 265 6000 245 5890 260 5840 p (t-test) 0.017 0.032 0.046 Min 0.0116 0.0116 0.0187 0.0116 0.0187 0.0116 Max 1160 53500 1160 53500 1160 53500 n (Patient) 57 259 46 270 41 275 sCr only Median 7.54 338 7.54 338 7.54 316 Average 169 2210 173 2190 180 2160 Stdev 314 6230 317 6200 325 6160 p (t-test) 0.0046 0.0054 0.0076 Min 0.0116 0.0116 0.0116 0.0116 0.0187 0.0116 Max 1300 53500 1300 53500 1300 53500 n (Patient) 76 239 74 241 70 245 UO only Median 118 248 104 275 103 237 Average 1880 1470 1750 1690 1460 1980 Stdev 6590 3060 6510 3970 5360 5640 p (t-test) 0.53 0.92 0.41 Min 0.0116 0.0116 0.0187 0.0116 0.0187 0.0116 Max 53500 19200 53500 30000 52500 53500 n (Patient) 193 121 172 142 155 159 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.72 0.55 0.71 0.71 0.56 0.72 0.70 0.57 SE 0.035 0.031 0.034 0.036 0.031 0.033 0.037 0.032 0.032 p Value 1.4E−8  2.1E−12 0.12 6.7E−9  1.9E−11 0.076 1.4E−9  3.8E−10 0.043 nCohort Recovered 57 76 193 46 74 172 41 70 155 nCohort Non- 259 239 121 270 241 142 275 245 159 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 79% 82% 79% 79% 81% 78% 79% 81% 79% Specificity 35% 39% 25% 37% 39% 24% 41% 39% 26% Cutoff Quartile 3 155 154 155 155 154 155 155 154 155 Sensitivity 56% 59% 55% 56% 58% 56% 55% 57% 55% Specificity 77% 76% 53% 83% 76% 55% 80% 74% 55% Cutoff Quartile 4 909 912 902 909 912 902 909 912 902 Sensitivity 30% 31% 29% 29% 31% 28% 28% 31% 28% Specificity 96% 95% 77% 96% 95% 77% 95% 94% 78% OR Quartile 2 2.05 2.89 1.27 2.19 2.81 1.16 2.71 2.65 1.33 p Value 0.023 2.3E−4 0.39 0.021 3.7E−4 0.59 0.0044 9.5E−4 0.29 Lower limit 1.10 1.64 0.735 1.13 1.59 0.682 1.36 1.49 0.785 of 95% CI Upper limit 3.82 5.08 2.20 4.26 4.95 1.96 5.38 4.71 2.25 of 95% CI OR Quartile 3 4.30 4.56 1.42 5.94 4.31 1.59 4.95 3.85 1.54 p Value 1.7E−5 4.3E−7 0.13 1.3E−5 1.2E−6 0.042 1.0E−4 8.2E−6 0.055 Lower limit 2.21 2.53 0.899 2.67 2.39 1.02 2.21 2.13 0.990 of 95% CI Upper limit 8.38 8.20 2.24 13.2 7.78 2.49 11.1 6.97 2.41 of 95% CI OR Quartile 4 11.6 8.23 1.38 8.78 7.91 1.34 7.58 7.28 1.40 p Value 8.1E−4 7.5E−5 0.22 0.0031 1.0E−4 0.26 0.0060 2.0E−4 0.19 Lower limit 2.77 2.90 0.822 2.08 2.78 0.802 1.79 2.56 0.841 of 95% CI Upper limit 48.9 23.4 2.31 37.1 22.5 2.23 32.2 20.7 2.35 of 95% CI

TABLE 18.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.1 373 18.3 355 9.44 346 Average 185 2330 176 2230 161 2170 Stdev 476 6380 492 6250 472 6150 p (t-test) 0.0016 0.0038 0.0065 Min 0.0116 0.0116 0.0187 0.0116 0.0187 0.0116 Max 3630 53500 3630 53500 3630 53500 n (Patient) 90 226 79 237 71 245 sCr only Median 18.3 403 18.3 394 18.3 373 Average 204 2480 208 2460 196 2410 Stdev 473 6590 477 6570 455 6500 p (t-test) 4.5E−4 5.7E−4 8.1E−4 Min 0.0116 0.0116 0.0116 0.0116 0.0187 0.0116 Max 3630 53500 3630 53500 3630 53500 n (Patient) 106 210 104 212 99 217 UO only Median 115 338 115 248 115 222 Average 1880 1410 2010 1260 1750 1690 Stdev 6360 3070 6630 2860 5640 5330 p (t-test) 0.48 0.24 0.93 Min 0.0116 0.0116 0.0151 0.0116 0.0151 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 211 103 193 121 177 137 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.73 0.55 0.73 0.73 0.53 0.73 0.73 0.54 SE 0.029 0.028 0.035 0.030 0.028 0.034 0.030 0.029 0.033 p Value 3.0E−14 0 0.18 3.1E−14 6.7E−16 0.33 8.2E−15 3.1E−15 0.23 nCohort Recovered 90 106 211 79 104 193 71 99 177 nCohort Non- 226 210 103 237 212 121 245 217 137 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 81% 84% 79% 81% 83% 78% 82% 83% 79% Specificity 36% 38% 24% 38% 38% 24% 41% 37% 25% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 61% 63% 57% 60% 62% 56% 58% 61% 55% Specificity 78% 75% 54% 80% 75% 54% 79% 75% 54% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 33% 35% 28% 32% 34% 26% 31% 34% 26% Specificity 96% 94% 76% 95% 94% 75% 96% 95% 76% OR Quartile 2 2.42 3.14 1.17 2.69 3.03 1.09 3.07 2.90 1.23 p Value 0.0015 3.1E−5 0.58 5.5E−4 5.2E−5 0.76 1.3E−4 1.1E−4 0.44 Lower limit 1.40 1.83 0.666 1.53 1.77 0.634 1.73 1.69 0.723 of 95% CI Upper limit 4.17 5.37 2.07 4.70 5.19 1.87 5.44 4.98 2.10 of 95% CI OR Quartile 3 5.49 5.21 1.55 5.89 4.95 1.50 5.23 4.69 1.40 p Value 3.3E−9 6.5E−10 0.072 1.0E−8 2.1E−9 0.083 2.0E−7 1.1E−8 0.14 Lower limit 3.12 3.08 0.962 3.21 2.93 0.949 2.80 2.76 0.896 of 95% CI Upper limit 9.65 8.79 2.49 10.8 8.36 2.37 9.77 7.96 2.19 of 95% CI OR Quartile 4 10.7 8.88 1.26 8.68 8.58 1.04 10.2 9.73 1.11 p Value 8.1E−6 9.0E−7 0.39 4.8E−5 1.3E−6 0.88 1.3E−4 2.2E−6 0.69 Lower limit 3.77 3.72 0.740 3.06 3.59 0.617 3.11 3.79 0.665 of 95% CI Upper limit 30.2 21.2 2.15 24.6 20.5 1.75 33.4 25.0 1.85 of 95% CI

TABLE 18.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 39.4 403 39.5 355 13.4 347 Average 365 2480 383 2400 386 2310 Stdev 1110 6700 1140 6600 1180 6460 p (t-test) 9.4E−4 0.0019 0.0040 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 7940 53500 7940 53500 7940 53500 n (Patient) 114 202 107 209 97 219 sCr only Median 36.6 423 39.4 403 36.6 366 Average 365 2620 373 2580 370 2530 Stdev 1060 6900 1070 6850 1080 6770 p (t-test) 3.0E−4 4.3E−4 6.4E−4 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 7940 53500 7940 53500 7940 53500 n (Patient) 127 189 124 192 119 197 UO only Median 118 338 118 237 115 222 Average 1870 1420 1950 1310 1740 1700 Stdev 6330 3100 6480 2970 5520 5490 p (t-test) 0.50 0.33 0.95 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 213 101 203 111 189 125 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.72 0.56 0.70 0.71 0.54 0.71 0.71 0.55 SE 0.029 0.028 0.035 0.030 0.029 0.034 0.030 0.029 0.033 p Value 8.5E−13 2.1E−14 0.10 1.4E−11 1.1E−12 0.19 4.2E−12 1.2E−12 0.15 nCohort Recovered 114 127 213 107 124 203 97 119 189 nCohort Non- 202 189 101 209 192 111 219 197 125 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 83% 85% 80% 83% 84% 80% 83% 84% 81% Specificity 34% 35% 25% 36% 35% 25% 38% 35% 26% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 62% 64% 57% 61% 63% 56% 60% 62% 55% Specificity 72% 71% 54% 72% 70% 53% 72% 70% 53% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 35% 36% 29% 33% 35% 26% 32% 35% 26% Specificity 92% 91% 77% 92% 91% 75% 92% 92% 76% OR Quartile 2 2.48 3.03 1.34 2.65 2.76 1.36 3.03 2.81 1.47 p Value 8.0E−4 5.3E−5 0.32 3.6E−4 1.9E−4 0.29 5.8E−5 1.5E−4 0.17 Lower limit of 1.46 1.77 0.751 1.55 1.62 0.772 1.77 1.65 0.849 95% CI Upper limit of 4.22 5.18 2.40 4.52 4.70 2.39 5.21 4.80 2.56 95% CI OR Quartile 3 4.25 4.33 1.55 4.06 4.01 1.44 3.86 3.75 1.41 p Value 1.2E−8 3.1E−9 0.071 5.6E−8 1.8E−8 0.13 3.5E−7 9.5E−8 0.13 Lower limit of 2.58 2.67 0.963 2.45 2.47 0.903 2.30 2.31 0.898 95% CI Upper limit of 6.99 7.03 2.50 6.72 6.50 2.29 6.49 6.09 2.23 95% CI OR Quartile 4 6.19 5.93 1.31 5.48 5.63 1.08 5.34 5.88 1.12 p Value 1.4E−6 3.7E−7 0.32 6.7E−6 7.9E−7 0.77 2.4E−5 1.0E−6 0.68 Lower limit of 2.95 2.98 0.769 2.61 2.84 0.637 2.45 2.89 0.664 95% CI Upper limit of 13.0 11.8 2.24 11.5 11.2 1.84 11.6 12.0 1.87 95% CI

TABLE 18.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 40.2 420 48.0 355 39.4 351 Average 430 2660 455 2540 464 2420 Stdev 1260 7000 1300 6860 1350 6690 p (t-test) 3.1E−4 8.7E−4 0.0022 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 8690 53500 8690 53500 8690 53500 n (Patient) 134 182 125 191 114 202 sCr only Median 40.2 436 44.1 420 40.2 406 Average 425 2740 431 2710 426 2680 Stdev 1240 7110 1250 7070 1250 7020 p (t-test) 1.7E−4 2.2E−4 2.7E−4 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 8690 53500 8690 53500 8690 53500 n (Patient) 140 176 138 178 135 181 UO only Median 118 248 123 223 118 222 Average 1860 1430 1930 1320 1720 1730 Stdev 6300 3130 6430 3010 5470 5580 p (t-test) 0.52 0.35 0.98 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 215 99 206 108 193 121 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.71 0.55 0.69 0.70 0.54 0.69 0.70 0.54 SE 0.029 0.029 0.035 0.030 0.029 0.034 0.030 0.029 0.034 p Value 2.3E−12 2.8E−13 0.13 2.5E−10 6.1E−12 0.25 2.2E−10 7.9E−12 0.19 nCohort Recovered 134 140 215 125 138 206 114 135 193 nCohort Non- 182 176 99 191 178 108 202 181 121 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 84% 85% 81% 83% 84% 81% 84% 83% 81% Specificity 33% 34% 25% 34% 33% 25% 36% 33% 26% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 64% 65% 58% 62% 64% 56% 61% 64% 55% Specificity 69% 69% 53% 69% 68% 53% 69% 68% 53% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 37% 38% 28% 35% 37% 26% 34% 37% 26% Specificity 91% 91% 76% 90% 91% 75% 90% 91% 76% OR Quartile 2 2.48 2.79 1.41 2.51 2.49 1.40 2.88 2.43 1.49 p Value 8.4E−4 1.9E−4 0.25 6.7E−4 8.3E−4 0.25 1.1E−4 0.0010 0.16 Lower limit of 1.45 1.63 0.785 1.48 1.46 0.790 1.69 1.43 0.854 95% CI Upper limit of 4.22 4.78 2.54 4.28 4.24 2.48 4.91 4.14 2.60 95% CI OR Quartile 3 4.08 4.01 1.56 3.64 3.81 1.40 3.51 3.73 1.42 p Value 7.1E−9 8.1E−9 0.069 1.2E−7 2.7E−8 0.15 4.5E−8 4.6E−8 0.13 Lower limit of 2.54 2.50 0.965 2.26 2.38 0.880 2.16 2.33 0.899 95% CI Upper limit of 6.57 6.43 2.52 5.88 6.10 2.24 5.73 5.98 2.24 95% CI OR Quartile 4 5.92 5.86 1.27 5.09 5.67 1.06 4.75 6.02 1.12 p Value 1.6E−7 8.5E−8 0.39 1.6E−6 1.5E−7 0.82 8.7E−6 1.2E−7 0.68 Lower limit of 3.05 3.07 0.740 2.62 2.97 0.624 2.39 3.10 0.663 95% CI Upper limit of 11.5 11.2 2.17 9.90 10.8 1.81 9.44 11.7 1.88 95% CI

TABLE 18.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine sample for the “revocered” and “non-recovered” cohorts where starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 72.4 406 72.6 351 72.6 351 Average 1650 1810 1680 1770 1680 1770 Stdev 6260 4140 6300 4100 6300 4100 p (t-test) 0.80 0.88 0.88 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 34200 53500 34200 53500 34200 n (Patient) 187 129 184 132 184 132 sCr only Median 53.2 531 54.2 492 55.3 488 Average 638 2810 648 2760 652 2740 Stdev 2630 7160 2660 7110 2670 7090 p (t-test) 3.9E−4 5.8E−4 6.6E−4 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 30000 53500 30000 53500 30000 53500 n (Patient) 159 157 156 160 155 161 UO only Median 116 300 118 248 116 242 Average 1840 1490 1860 1450 1880 1430 Stdev 6370 3110 6410 3070 6450 3030 p (t-test) 0.60 0.53 0.49 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 210 104 207 107 204 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.70 0.57 0.63 0.69 0.56 0.63 0.69 0.57 SE 0.032 0.029 0.035 0.032 0.029 0.035 0.032 0.030 0.034 p Value 3.1E−5 3.6E−12 0.038 7.9E−5 4.5E−11 0.068 7.9E−5 1.3E−10 0.054 nCohort Recovered 187 159 210 184 156 207 184 155 204 nCohort Non- 129 157 104 132 160 107 132 161 110 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 85% 85% 82% 85% 84% 81% 85% 84% 82% Specificity 29% 32% 26% 29% 31% 26% 29% 31% 26% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 64% 66% 60% 64% 66% 58% 64% 65% 58% Specificity 60% 66% 55% 60% 66% 54% 60% 66% 54% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 34% 39% 31% 33% 38% 30% 33% 38% 30% Specificity 81% 89% 78% 81% 88% 77% 81% 88% 77% OR Quartile 2 2.41 2.75 1.55 2.33 2.47 1.50 2.33 2.33 1.58 p Value 0.0029 3.4E−4 0.14 0.0038 0.0011 0.17 0.0038 0.0022 0.12 Lower limit of 95% 1.35 1.58 0.862 1.31 1.43 0.840 1.31 1.36 0.887 CI Upper limit of 95% CI 4.31 4.79 2.78 4.12 4.26 2.67 4.12 4.00 2.81 OR Quartile 3 2.69 3.82 1.79 2.60 3.71 1.62 2.60 3.61 1.66 p Value 2.8E−5 1.8E−8 0.017 4.8E−5 3.3E−8 0.044 4.8E−5 5.9E−8 0.034 Lower limit of 95% 1.69 2.39 1.11 1.64 2.33 1.01 1.64 2.27 1.04 CI Upper limit of 95% CI 4.29 6.08 2.88 4.13 5.91 2.60 4.13 5.74 2.65 OR Quartile 4 2.25 4.98 1.54 2.13 4.72 1.45 2.13 4.64 1.47 p Value 0.0021 8.1E−8 0.11 0.0041 2.0E−7 0.16 0.0041 2.8E−7 0.15 Lower limit of 95% 1.34 2.77 0.909 1.27 2.63 0.858 1.27 2.59 0.872 CI Upper limit of 95% CI 3.77 8.95 2.61 3.57 8.49 2.46 3.57 8.34 2.48

TABLE 18.6 Comparison of marker levels and the area under the ROC curve (AUC) in urine sample for the “revocered” and “non-recovered” cohorts where starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 2350 4630 2350 4520 2350 4520 Average 2780 5040 2970 4940 2740 4940 Stdev 2070 3020 2160 3030 1870 3030 p (t-test) 4.6E−4 0.0047 0.0024 Min 62.8 413 62.8 276 62.8 276 Max 8250 21200 8250 21200 8050 21200 n (Patient) 25 126 21 130 19 132 sCr only Median 3060 4590 3080 4560 3240 4520 Average 3540 4950 3570 4930 3720 4870 Stdev 2350 3090 2390 3080 2410 3090 p (t-test) 0.023 0.031 0.076 Min 842 62.8 842 62.8 842 62.8 Max 10700 21200 10700 21200 10700 21200 n (Patient) 29 121 28 122 26 124 UO only Median 4050 4670 3990 4570 3790 4710 Average 4550 4910 4490 4940 4200 5210 Stdev 2870 3190 2730 3300 2440 3430 p (t-test) 0.47 0.37 0.040 Min 62.8 513 62.8 276 62.8 276 Max 13800 21200 13800 21200 11800 21200 n (Patient) 91 59 84 66 77 73 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.76 0.66 0.54 0.73 0.65 0.54 0.75 0.63 0.59 SE 0.045 0.052 0.048 0.052 0.053 0.048 0.051 0.056 0.047 p Value 7.1E−9 0.0020 0.41 1.3E−5 0.0037 0.41 7.7E−7 0.022 0.064 nCohort Recovered 25 29 91 21 28 84 19 26 77 nCohort Non- 126 121 59 130 122 66 132 124 73 recovered Cutoff Quartile 2 2740 2730 2740 2740 2730 2740 2740 2730 2740 Sensitivity 81% 79% 80% 79% 78% 77% 79% 77% 79% Specificity 56% 41% 29% 52% 39% 27% 53% 35% 30% Cutoff Quartile 3 4230 4260 4260 4230 4260 4260 4230 4260 4260 Sensitivity 58% 56% 54% 56% 56% 55% 56% 55% 58% Specificity 88% 76% 53% 86% 75% 54% 89% 73% 57% Cutoff Quartile 4 6010 6060 6060 6010 6060 6060 6010 6060 6060 Sensitivity 29% 29% 20% 28% 29% 23% 28% 28% 26% Specificity 92% 90% 71% 90% 89% 73% 95% 88% 75% OR Quartile 2 5.41 2.58 1.57 4.20 2.28 1.28 4.13 1.73 1.65 p Value 2.6E−4 0.030 0.26 0.0033 0.064 0.52 0.0051 0.23 0.19 Lower limit of 95% CI 2.19 1.09 0.718 1.61 0.953 0.606 1.53 0.699 0.779 Upper limit of 95% CI 13.4 6.08 3.42 10.9 5.44 2.71 11.1 4.30 3.48 OR Quartile 3 10.1 4.03 1.32 7.68 3.78 1.38 10.8 3.30 1.81 p Value 3.1E−4 0.0031 0.40 0.0017 0.0050 0.32 0.0019 0.013 0.073 Lower limit of 95% CI 2.87 1.60 0.686 2.16 1.49 0.725 2.41 1.29 0.946 Upper limit of 95% CI 35.5 10.2 2.55 27.4 9.55 2.64 48.8 8.40 3.45 OR Quartile 4 4.60 3.53 0.638 3.64 3.35 0.780 7.01 3.01 1.07 p Value 0.046 0.050 0.26 0.093 0.060 0.52 0.063 0.087 0.85 Lower limit of 95% CI 1.03 1.00 0.293 0.806 0.951 0.369 0.903 0.851 0.514 Upper limit of 95% CI 20.5 12.4 1.39 16.4 11.8 1.65 54.4 10.7 2.24

TABLE 18.7 Comparison of marker levels and the area under the ROC curve (AUC) in urine sample for the “revocered” and “non-recovered” cohorts where starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3060 4710 3080 4670 3080 4590 Average 3490 5090 3580 5010 3480 4990 Stdev 2270 3120 2360 3100 2330 3080 p (t-test) 0.0035 0.012 0.011 Min 62.8 276 62.8 276 62.8 276 Max 11800 21200 11800 21200 11800 21200 n (Patient) 40 111 36 115 32 119 sCr only Median 3500 4700 3620 4680 3800 4530 Average 3860 5000 3890 4980 4030 4900 Stdev 2390 3160 2410 3160 2440 3150 p (t-test) 0.034 0.043 0.12 Min 842 62.8 842 62.8 842 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 44 107 43 108 40 111 UO only Median 4090 4690 4060 4670 3920 4710 Average 4590 4890 4520 4970 4260 5250 Stdev 2740 3460 2660 3480 2400 3570 p (t-test) 0.56 0.38 0.045 Min 62.8 276 62.8 276 62.8 276 Max 13800 21200 13800 21200 11800 21200 n (Patient) 100 50 93 57 85 65 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.68 0.62 0.52 0.66 0.62 0.53 0.67 0.59 0.58 SE 0.046 0.048 0.050 0.049 0.049 0.049 0.050 0.051 0.047 p Value 1.3E−4 0.010 0.66 0.0014 0.016 0.54 8.8E−4 0.079 0.10 nCohort Recovered 40 44 100 36 43 93 32 40 85 nCohort Non- 111 107 50 115 108 57 119 111 65 recovered Cutoff Quartile 2 2740 2740 2740 2740 2740 2740 2740 2740 2740 Sensitivity 78% 77% 74% 77% 76% 74% 77% 75% 77% Specificity 35% 30% 25% 33% 28% 25% 34% 25% 27% Cutoff Quartile 3 4230 4230 4260 4230 4230 4260 4230 4230 4260 Sensitivity 59% 58% 56% 57% 57% 56% 57% 56% 58% Specificity 75% 68% 53% 72% 67% 54% 75% 65% 56% Cutoff Quartile 4 6010 6010 6060 6010 6010 6060 6010 6010 6060 Sensitivity 31% 31% 24% 30% 31% 25% 29% 30% 28% Specificity 90% 89% 74% 89% 88% 74% 91% 88% 76% OR Quartile 2 1.95 1.38 0.949 1.71 1.22 0.920 1.78 0.988 1.24 p Value 0.098 0.43 0.89 0.20 0.62 0.83 0.18 0.98 0.58 Lower limit of 95% CI 0.885 0.626 0.436 0.754 0.549 0.433 0.766 0.429 0.584 Upper limit of 95% CI 4.31 3.02 2.06 3.88 2.71 1.96 4.16 2.28 2.62 OR Quartile 3 4.40 2.95 1.44 3.50 2.79 1.49 4.00 2.35 1.83 p Value 3.4E−4 0.0042 0.30 0.0027 0.0068 0.24 0.0020 0.026 0.071 Lower limit of 95% CI 1.96 1.41 0.725 1.55 1.33 0.767 1.66 1.11 0.950 Upper limit of 95% CI 9.89 6.20 2.84 7.93 5.87 2.89 9.63 4.97 3.51 OR Quartile 4 3.97 3.48 0.899 3.36 3.34 0.936 4.03 2.96 1.24 p Value 0.015 0.016 0.79 0.033 0.020 0.86 0.029 0.037 0.56 Lower limit of 95% CI 1.31 1.26 0.409 1.10 1.21 0.437 1.15 1.07 0.594 Upper limit of 95% CI 12.0 9.62 1.98 10.2 9.26 2.00 14.1 8.23 2.61

TABLE 18.8 Comparison of marker levels and the area under the ROC curve (AUC) in urine sample for the “revocered” and “non-recovered” cohorts where starts within 72hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3500 4670 3500 4530 3500 4530 Average 3980 5010 3930 4990 3900 4940 Stdev 2670 3100 2540 3130 2570 3100 p (t-test) 0.047 0.046 0.059 Min 62.8 276 62.8 276 62.8 276 Max 11800 21200 11800 21200 11800 21200 n (Patient) 50 101 46 105 40 111 sCr only Median 3860 4530 3930 4500 3940 4480 Average 4130 4960 4230 4900 4350 4820 Stdev 2520 3210 2520 3210 2530 3200 p (t-test) 0.10 0.19 0.38 Min 703 62.8 842 62.8 842 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 54 97 52 99 49 102 UO only Median 3930 4790 3930 4790 3770 4790 Average 4450 5150 4410 5160 4270 5250 Stdev 2680 3490 2640 380 2590 3400 p (t-test) 017 0.14 0.048 Min 62.8 276 62.8 276 62.8 276 Max 13800 21200 13800 21200 13800 21200 n (Patient) 98 52 94 56 86 64 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.62 0.58 0.56 0.61 0.56 0.57 0.61 0.54 0.59 SE 0.047 0.048 0.050 0.048 0.048 0.049 0.050 0.050 0.047 p Value 0.013 0.078 0.19 0.020 0.19 0.16 0.023 0.45 0.045 nCohort Recovered 50 54 98 46 52 94 40 49 86 nCohort Non- 101 91 52 105 99 56 111 102 64 recovered Cutoff Quartile 2 2740 2740 2740 2740 2740 2740 2740 2740 2740 Sensitivity 78% 76% 75% 77% 75% 75% 77% 74% 78% Specificity 32% 28% 26% 30% 25% 26% 30% 22% 28% Cutoff Quartile 3 4230 4230 4260 4230 4230 4260 4230 4230 4260 Sensitivity 58% 57% 62% 57% 56% 61% 57% 54% 61% Specificity 66% 61% 56% 65% 60% 56% 68% 57% 58% Cutoff Quartile 4 6010 6010 6060 6010 6010 6060 6010 6010 6060 Sensitivity 29% 30% 27% 29% 29% 27% 28% 28% 28% Specificity 82% 83% 76% 83% 83% 76% 82% 82% 77% OR Quartile 2 1.69 1.24 1.03 1.48 0.987 1.03 1.40 0.804 1.38 p Value 0.18 0.58 0.95 0.32 0.97 0.94 0.41 0.59 0.40 Lower limit of 95% CI 0.791 0.580 0.473 0.680 0.455 0.480 0.626 0.360 0.648 Upper limit of 95% CI 3.61 2.64 2.23 3.21 2.14 2.20 3.14 1.78 2.95 OR Quartile 3 2.73 2.03 2.05 2.50 1.85 2.00 2.73 1.56 2.17 p Value 0.0054 0.037 0.041 0.013 0.078 0.044 0.0098 0.20 0.022 Lower limit of 95% CI 1.35 1.04 1.03 1.22 0.934 1.02 1.27 0.784 1.12 Upper limit of 95% CI 5.53 4.03 4.07 5.13 3.65 3.92 5.83 3.10 4.19 OR Quartile 4 1.83 2.13 1.14 1.90 1.98 1.13 1.83 1.77 1.29 p Value 0.16 0.176 0.74 0.15 0.11 0.75 0.20 0.19 0.50 Lower limit of 95% CI 0.792 0.923 0.528 0.794 0.856 0.530 0.732 0.761 0.616 Upper limit of 95% CI 4.25 4.93 2.44 4.54 4.58 2.40 4.56 4.10 2.71

TABLE 18.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3650 4700 3620 4600 3540 4630 Average 4130 5000 4100 4970 3970 4980 Stdev 2690 3140 2620 3150 2610 3120 p (t-test) 0.084 0.087 0.053 Min 62.8 276 62.8 276 62.8 276 Max 11800 21200 11800 21200 11800 21200 n (Patient) 58 93 53 98 47 104 sCr only Median 3860 4530 3920 4520 3930 4490 Average 4250 4940 4310 4890 4390 4830 Stdev 2600 3220 2580 3230 2590 3220 p (t-test) 0.17 0.25 0.38 Min 703 62.8 842 62.8 842 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 60 91 59 92 57 94 UO only Median 3930 4820 3920 4790 3770 4790 Average 4430 5150 4400 5150 4270 5250 Stdev 2710 3420 2660 3420 2590 3400 p (t-test) 0.16 0.14 0.048 Min 62.8 276 62.8 276 62.8 276 Max 13800 21200 13800 21200 13800 21200 n (Patient) 96 54 92 58 86 64 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.60 0.57 0.57 0.59 0.56 0.57 0.61 0.54 0.60 SE 0.047 0.047 0.049 0.047 0.048 0.048 0.048 0.048 0.047 p Value 0.034 0.14 0.13 0.048 0.22 0.14 0.017 0.39 0.044 nCohort 58 60 96 53 59 92 47 57 86 Recovered nCohort Non- 93 91 54 98 92 58 104 94 64 recovered Cutoff Quartile 2 2740 2740 2740 2740 2740 2740 2740 2740 2740 Sensitivity 78% 77% 76% 78% 76% 76% 78% 76% 78% Specificity 31% 28% 26% 30% 27% 26% 32% 26% 28% Cutoff Quartile 3 4230 4230 4260 4230 4230 4260 4230 4230 4260 Sensitivity 59% 57% 61% 58% 57% 60% 59% 55% 61% Specificity 64% 60% 56% 64% 59% 57% 68% 58% 58% Cutoff Quartile 4 6010 6010 6060 6010 6010 6060 6010 6010 6060 Sensitivity 28% 29% 26% 28% 28% 26% 28% 28% 28% Specificity 79% 80% 75% 79% 80% 75% 81% 79% 77% OR Quartile 2 1.64 1.32 1.11 1.49 1.18 1.11 1.65 1.10 1.38 p Value 0.19 0.47 0.79 0.30 0.66 0.79 0.20 0.80 0.40 Lower limit of 0.780 0.626 0.513 0.703 0.561 0.519 0.765 0.519 0.648 95% CI Upper limit of 3.46 2.77 2.40 3.18 2.50 2.37 3.56 2.34 2.95 95% CI OR Quartile 3 2.55 2.00 2.02 2.49 1.90 1.98 3.03 1.70 2.17 p Value 0.0067 0.040 0.043 0.0096 0.059 0.045 0.0028 0.12 0.022 Lower limit of 1.30 1.03 1.02 1.25 0.977 1.01 1.46 0.876 1.12 95% CI Upper limit of 5.02 3.88 3.99 4.96 3.68 3.86 6.26 3.31 4.19 95% CI OR Quartile 4 1.49 1.60 1.05 1.45 1.54 1.05 1.63 1.43 1.29 p Value 0.32 0.24 0.90 0.36 0.28 0.91 0.25 0.37 0.50 Lower limit of 0.682 0.734 0.489 0.654 0.708 0.492 0.702 0.657 0.616 95% CI Upper limit of 3.25 3.49 2.25 3.23 3.36 2.22 3.80 3.13 2.71 95% CI

TABLE 18.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 4170 4480 4150 4470 4150 4470 Average 4620 4720 4620 4720 4620 4720 Stdev 2740 3290 2760 3260 2760 3260 p (t-test) 0.83 0.84 0.84 Min 703 62.8 703 62.8 703 62.8 Max 13800 21200 13800 21200 13800 21200 n (Patient) 81 70 80 71 80 71 sCr only Median 3920 4520 3920 4520 3920 4520 Average 4320 4940 4360 4890 4360 4890 Stdev 2570 3290 2570 3280 2570 3280 p (t-test) 0.21 0.28 0.28 Min 703 62.8 842 62.8 842 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 67 84 65 86 65 86 UO only Median 3940 4710 3940 4670 3940 4670 Average 4530 4970 4570 4890 4560 4900 Stdev 2690 3460 2690 3440 2720 3390 p (t-test) 0.39 0.52 0.49 Min 413 62.8 413 62.8 413 62.8 Max 13800 21200 13800 21200 13800 21200 n (Patient) 95 55 93 57 91 59 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.50 0.56 0.54 0.51 0.55 0.53 0.51 0.55 0.53 SE 0.047 0.047 0.049 0.047 0.047 0.049 0.047 0.047 0.049 p Value 0.92 0.23 0.44 0.91 0.31 0.59 0.91 0.31 0.52 nCohort Recovered 81 67 95 80 65 93 80 65 91 nCohort Non- 70 84 55 71 86 57 71 86 59 recovered Cutoff Quartile 2 2740 2740 2740 2740 2740 2740 2740 2740 2740 Sensitivity 73% 77% 75% 73% 77% 74% 73% 77% 75% Specificity 23% 28% 25% 24% 28% 25% 24% 28% 25% Cutoff Quartile 3 4230 4230 4260 4230 4230 4260 4230 4230 4260 Sensitivity 53% 57% 56% 54% 57% 56% 54% 57% 56% Specificity 52% 58% 54% 52% 58% 54% 52% 58% 54% Cutoff Quartile 4 6010 6010 6060 6010 6010 6060 6010 6010 6060 Sensitivity 24% 26% 24% 24% 26% 23% 24% 26% 24% Specificity 74% 76% 74% 74% 75% 73% 74% 75% 74% OR Quartile 2 0.823 1.35 0.990 0.852 1.26 0.920 0.852 1.26 0.992 p Value 0.60 0.42 0.98 0.67 0.53 0.83 0.67 0.53 0.98 Lower limit 0.394 0.648 0.462 0.408 0.604 0.433 0.408 0.604 0.467 of 95% CI Upper limit 1.72 2.83 2.12 1.78 2.65 1.96 1.78 2.65 2.11 of 95% CI OR Quartile 3 1.21 1.86 1.50 1.27 1.86 1.49 1.27 1.86 1.48 p Value 0.56 0.062 0.24 0.46 0.061 0.24 0.46 0.061 0.24 Lower limit 0.636 0.970 0.767 0.671 0.971 0.767 0.671 0.971 0.766 of 95% CI Upper limit 2.29 3.56 2.92 2.41 3.58 2.89 2.41 3.58 2.86 of 95% CI OR Quartile 4 0.916 1.13 0.867 0.884 1.05 0.804 0.884 1.05 0.869 p Value 0.82 0.75 0.72 0.74 0.89 0.58 0.74 0.89 0.72 Lower limit 0.438 0.538 0.401 0.423 0.500 0.372 0.423 0.500 0.406 of 95% CI Upper limit 1.92 2.38 1.87 1.85 2.21 1.74 1.85 2.21 1.86 of 95% CI

Example 19. Use of C—C Motif Chemokine 15 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 15 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 19.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.1 401 36.6 366 37.9 363 Average 820 2170 852 2130 869 2110 Stdev 4380 5930 4460 5880 4530 5840 p (t-test) 0.038 0.051 0.062 Min 0.0116 0.0116 0.0187 0.0116 0.0187 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 107 209 103 213 100 216 sCr only Median 22.1 414 22.1 403 24.3 401 Average 754 2360 765 2330 771 2320 Stdev 4060 6190 4090 6160 4110 6150 p (t-test) 0.011 0.013 0.015 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 125 190 123 192 122 193 UO only Median 110 347 115 341 118 261 Average 1780 1580 1880 1420 1670 1810 Stdev 6210 3290 6400 3100 5420 5670 p (t-test) 0.77 0.49 0.83 Min 0.0116 0.0116 0.0187 0.0116 0.0187 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 221 93 207 107 197 117 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.72 0.72 0.57 0.70 0.72 0.55 0.70 0.72 0.55 SE 0.029 0.028 0.036 0.030 0.028 0.035 0.030 0.028 0.034 p Value 1.2E−14 1.1E−15 0.038 5.3E−12 2.7E−15 0.13 1.5E−11 1.5E−14 0.17 nCohort 107 125 221 103 123 207 100 122 197 Recovered nCohort Non- 209 190 93 213 192 107 216 193 117 recovered Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 83% 85% 81% 82% 85% 79% 82% 84% 78% Specificity 36% 36% 25% 35% 37% 24% 35% 36% 24% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 63% 65% 60% 62% 65% 58% 61% 64% 56% Specificity 75% 72% 54% 74% 72% 54% 74% 72% 54% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 34% 36% 30% 33% 36% 27% 33% 36% 28% Specificity 93% 92% 77% 92% 92% 76% 92% 92% 77% OR Quartile 2 2.85 3.12 1.38 2.47 3.24 1.16 2.44 3.06 1.10 p Value 1.3E−4 3.4E−5 0.29 9.2E−4 1.9E−5 0.60 0.0011 4.3E−5 0.74 Lower limit 1.67 1.82 0.759 1.45 1.89 0.664 1.43 1.79 0.636 of 95% CI Upper limit 4.87 5.35 2.51 4.23 5.56 2.04 4.18 5.24 1.89 of 95% CI OR Quartile 3 4.98 4.72 1.80 4.50 4.77 1.62 4.47 4.65 1.51 p Value 1.3E−9 5.8E−10 0.020 1.3E−8 5.4E−10 0.044 2.1E−8 1.0E−9 0.081 Lower limit 2.96 2.89 1.10 2.68 2.91 1.01 2.65 2.84 0.951 of 95% CI Upper limit 8.36 7.71 2.94 7.55 7.82 2.60 7.55 7.62 2.39 of 95% CI OR Quartile 4 6.37 6.56 1.44 5.94 6.34 1.17 5.63 6.23 1.29 p Value 2.9E−6 2.1E−7 0.19 6.8E−6 3.5E−7 0.57 1.3E−5 4.5E−7 0.34 Lower limit 2.93 3.22 0.835 2.73 3.11 0.686 2.59 3.06 0.766 of 95% CI Upper limit 13.8 13.3 2.47 12.9 12.9 1.99 12.2 12.7 2.17 of 95% CI

TABLE 19.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 48.0 434 49.4 434 53.2 423 Average 874 2340 888 2320 908 2270 Stdev 4000 6310 4030 6280 4090 6220 p (t-test) 0.018 0.022 0.029 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 135 181 133 183 129 187 sCr only Median 48.7 436 49.4 434 51.3 434 Average 836 2490 841 2480 847 2460 Stdev 3820 6530 3840 6510 3850 6490 p (t-test) 0.0072 0.0080 0.0089 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 148 168 147 169 146 170 UO Only Median 106 354 110 347 113 300 Average 1780 1580 1820 1500 1650 1850 Stdev 6170 3270 6250 3180 5320 5850 p (t-test) 0.78 0.63 0.76 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 225 89 219 95 206 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.70 0.70 0.58 0.69 0.70 0.57 0.68 0.69 0.55 SE 0.029 0.029 0.036 0.029 0.029 0.036 0.030 0.029 0.034 p Value 1.4E−11 3.5E−12 0.021 1.3E−10 8.5E−12 0.051 1.1E−9 3.3E−11 0.12 nCohort 135 148 225 133 147 219 129 146 206 Recovered nCohort Non- 181 168 89 183 169 95 187 170 108 recovered Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 84% 86% 81% 84% 86% 80% 83% 85% 79% Specificity 33% 34% 25% 33% 34% 25% 33% 34% 24% Cutoff 155 155 155 155 155 155 155 155 155 Quartile 3 Sensitivity 64% 65% 62% 63% 64% 60% 63% 64% 57% Specificity 69% 67% 55% 68% 67% 54% 68% 66% 54% Cutoff 909 909 902 909 909 902 909 909 902 Quartile 4 Sensitivity 35% 37% 30% 35% 37% 28% 34% 36% 28% Specificity 89% 89% 77% 89% 88% 76% 88% 88% 76% OR Quartile 2 2.62 3.06 1.40 2.52 3.11 1.31 2.34 2.93 1.18 p Value 4.1E−4 6.7E−5 0.28 6.7E−4 5.2E−5 0.37 0.0017 1.1E−4 0.55 Lower limit of 1.54 1.77 0.763 1.48 1.80 0.726 1.38 1.70 0.677 95% CI Upper limit of 4.47 5.31 2.58 4.29 5.40 2.36 3.97 5.06 2.07 95% CI OR Quartile 3 3.95 3.73 1.95 3.75 3.63 1.78 3.59 3.54 1.57 p Value 1.4E−8 3.1E−8 0.0090 4.4E−8 5.6E−8 0.020 1.3E−7 1.0E−7 0.058 Lower limit of 2.46 2.34 1.18 2.34 2.28 1.09 2.23 2.22 0.985 95% CI Upper limit of 6.35 5.95 3.22 6.02 5.79 2.91 5.77 5.63 2.52 95% CI OR Quartile 4 4.38 4.51 1.45 4.23 4.43 1.28 3.95 4.36 1.23 p Value 2.8E−6 6.7E−7 0.18 4.6E−6 9.2E−7 0.38 1.3E−5 1.2E−6 0.44 Lower limit of 2.36 2.49 0.838 2.28 2.45 0.740 2.13 2.40 0.726 95% CI Upper limit of 8.11 8.17 2.51 7.84 8.03 2.20 7.33 7.89 2.09 95% CI

TABLE 19.3 Comparison of marker level and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum sreatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 49.4 537 49.4 497 53.2 437 Average 877 2500 888 2470 912 2420 Stdev 3820 6600 3850 6560 3900 6490 p (t-test) 0.0082 0.010 0.015 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 153 163 151 165 147 169 sCr only Median 53.2 517 53.2 497 54.2 492 Average 864 2590 870 2570 875 2560 Stdev 3740 6730 3750 6710 3760 6690 p (t-test) 0.0050 0.0056 0.0062 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 160 156 159 157 158 158 UO only Median 115 354 113 351 115 261 Average 1790 1550 1800 1530 1640 1900 Stdev 6200 3230 6210 3220 5260 5980 p (t-test) 0.72 0.69 0.69 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 223 91 222 92 211 103 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.70 0.58 0.70 0.69 0.58 0.68 0.69 0.56 SE 0.029 0.029 0.036 0.029 0.029 0.036 0.030 0.030 0.035 p Value 9.2E−12 2.0E−11 0.031 2.3E−11 4.7E−11 0.032 9.8E−10 1.7E−10 0.081 nCohort Recovered 153 160 223 151 159 222 147 158 211 nCohort Non- 163 156 91 165 157 92 169 158 103 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 84% 85% 80% 84% 85% 80% 83% 85% 80% Specificity 31% 32% 25% 32% 32% 25% 31% 32% 25% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 66% 66% 62% 65% 66% 61% 64% 65% 57% Specificity 67% 66% 55% 67% 65% 55% 66% 65% 54% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 37% 38% 31% 37% 38% 30% 36% 37% 28% Specificity 88% 88% 77% 88% 87% 77% 88% 87% 76% OR Quartile 2 2.41 2.71 1.33 2.49 2.75 1.35 2.29 2.58 1.28 Upper limit of 95% CI p Value 0.0014 4.3E−4 0.35 9.4E−4 3.4E−4 0.32 0.0023 6.9E−4 0.40 Lower limit of 95% 1.40 1.56 0.729 1.45 1.58 0.744 1.34 1.49 0.720 CI 4.14 4.71 2.42 4.28 4.79 2.46 3.92 4.47 2.26 OR Quartile 3 4.05 3.71 1.93 3.83 3.61 1.86 3.43 3.51 1.55 Upper limit of 95% CI p Value 5.0E−9 3.3E−8 0.0095 1.7E−8 6.0E−8 0.014 1.8E−7 1.1E−7 0.072 Lower limit of 95% 2.53 2.33 1.17 2.40 2.27 1.14 2.16 2.21 0.962 CI 6.46 5.91 3.18 6.10 5.74 3.06 5.46 5.57 2.49 OR Quartile 4 4.49 4.26 1.50 4.33 4.18 1.47 4.05 4.11 1.26 Upper limit of 95% CI p Value 5.0E−7 6.1E−7 0.14 9.0E−7 8.3E−7 0.17 2.8E−6 1.1E−6 0.39 Lower limit of 95% 2.50 2.41 0.870 2.41 2.37 0.852 2.26 2.33 0.740 CI 8.05 7.52 2.58 7.78 7.39 2.53 7.26 7.26 2.15

TABLE 19.4 Comparison of marker level and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum sreatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 53.2 555 54.2 544 57.7 492 Average 856 2680 871 2630 893 2560 Stdev 3670 6870 3700 6810 3740 6740 p (t-test) 0.0030 0.0043 0.0067 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 167 149 164 152 160 156 sCR only Median 55.3 555 55.7 553 56.1 550 Average 847 2740 852 2720 857 2700 Stdev 3620 6950 3630 6930 3640 6910 p (t-test) 0.0021 0.0024 0.0027 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 171 145 170 146 169 147 UO only Median 116 340 118 338 118 248 Average 1810 1510 1820 1500 1630 1920 Stdev 6210 3220 6220 3210 5240 6040 p (t-test) 0.66 0.64 0.67 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 52500 53500 n (Patient) 222 92 221 93 213 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.70 0.57 0.69 0.70 0.57 0.68 0.69 0.56 SE 0.030 0.030 0.036 0.030 0.030 0.036 0.030 0.030 0.035 p Value 1.9E−11 9.6E−12 0.043 1.6E−10 2.4E−11 0.062 5.7E−9 9.1E−11 0.11 nCohort Recovered 167 171 222 164 170 221 160 169 213 nCohort Non- 149 145 92 152 146 93 156 147 101 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 85% 86% 82% 84% 86% 81% 83% 85% 80% Specificity 31% 31% 25% 30% 31% 25% 30% 31% 25% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 68% 68% 61% 66% 67% 60% 65% 67% 57% Specificity 66% 65% 55% 65% 65% 54% 64% 64% 54% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 38% 39% 29% 38% 38% 29% 37% 38% 28% Specificity 87% 87% 77% 87% 86% 76% 86% 86% 76% OR Quartile 2 2.41 2.65 1.49 2.34 2.70 1.38 2.14 2.53 1.34 Upper limit of 95% CI p Value 0.0018 7.1E−4 0.20 0.0024 5.8E−4 0.29 0.0057 0.0012 0.32 Lower limit of 95% 1.39 1.51 0.811 1.35 1.53 0.759 1.25 1.44 0.751 CI 4.19 4.67 2.73 4.05 4.74 2.51 3.68 4.41 2.40 OR Quartile 3 4.06 3.86 1.86 3.72 3.74 1.80 3.32 3.63 1.55 Upper limit of 95% CI p Value 4.9E−9 1.6E−8 0.014 3.2E−8 3.1E−8 0.020 3.4E−7 5.6E−8 0.071 Lower limit of 95% 2.54 2.41 1.14 2.33 2.35 1.10 2.09 2.28 0.963 CI 6.49 6.16 3.06 5.92 5.97 2.94 5.26 5.79 2.50 OR Quartile 4 4.08 4.05 1.36 3.87 3.98 1.33 3.61 3.91 1.22 Upper limit of 95% CI p Value 7.4E−7 6.9E−7 0.27 1.8E−6 9.4E−7 0.31 5.9E−6 1.3E−6 0.47 Lower limit of 95% 2.34 2.33 0.787 2.22 2.29 0.771 2.07 2.25 0.712 CI 7.13 7.03 2.34 6.76 6.90 2.29 6.29 6.78 2.09

TABLE 19.5 Comparison of marker level and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum sreatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 81.2 447 81.2 380 81.4 354 Average 1830 1510 1850 1480 1860 1470 Stdev 6490 2980 6520 2960 6530 2950 p (t-test) 0.61 0.56 0.54 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 202 114 200 116 199 117 sCr only Median 65.1 553 65.1 553 65.1 553 Average 943 2790 943 2790 943 2790 Stdev 3600 7230 3600 7230 3600 7230 p (t-test) 0.0029 0.0029 0.0029 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 184 132 184 132 184 132 UO only Median 108 344 110 341 110 338 Average 1790 1570 1800 1550 1820 1520 Stdev 6280 3180 6300 3170 6320 3140 p (t-test) 0.73 0.70 0.65 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 216 98 215 99 213 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.61 0.67 0.59 0.61 0.67 0.58 0.61 0.67 0.58 SE 0.033 0.031 0.035 0.033 0.031 0.035 0.033 0.031 0.035 p Value 6.4E−4 2.4E−8 0.016 0.0012 2.4E−8 0.024 0.0016 2.4E−8 0.031 nCohort Recovered 202 184 216 200 184 215 199 184 213 nCohort Non- 114 132 98 116 132 99 117 132 101 recovered Cutoff Quartile 2 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Sensitivity 84% 84% 82% 84% 84% 81% 84% 84% 81% Specificity 28% 29% 25% 28% 29% 25% 28% 29% 25% Cutoff Quartile 3 155 155 155 155 155 155 155 155 155 Sensitivity 65% 67% 62% 64% 67% 62% 63% 67% 60% Specificity 58% 62% 56% 58% 62% 55% 58% 62% 55% Cutoff Quartile 4 909 909 902 909 909 902 909 909 902 Sensitivity 33% 39% 32% 33% 39% 31% 32% 39% 31% Specificity 80% 85% 78% 80% 85% 78% 79% 85% 77% OR Quartile 2 2.05 2.14 1.52 1.94 2.14 1.41 1.97 2.14 1.47 Upper limit of 95% CI p Value 0.017 0.0084 0.17 0.026 0.0084 0.25 0.022 0.0084 0.20 Lower limit of 95% 1.13 1.22 0.837 1.08 1.22 0.785 1.10 1.22 0.815 CI 3.69 3.76 2.76 3.46 3.76 2.54 3.52 3.76 2.64 OR Quartile 3 2.60 3.26 2.06 2.43 3.26 1.99 2.36 3.26 1.86 Upper limit of 95% CI p Value 8.3E−5 7.8E−7 0.0037 2.2E−4 7.8E−7 0.0055 3.5E−4 7.8E−7 0.012 Lower limit of 95% 1.62 2.04 1.26 1.52 2.04 1.22 1.47 2.04 1.15 CI 4.18 5.20 3.36 3.90 5.20 3.24 3.77 5.20 3.01 OR Quartile 4 1.96 3.51 1.62 1.89 3.51 1.59 1.85 3.51 1.52 Upper limit of 95% CI p Value 0.011 4.0E−6 0.076 0.016 4.0E−6 0.089 0.019 4.0E−6 0.12 Lower limit of 95% 1.17 2.06 0.950 1.13 2.06 0.931 1.10 2.06 0.895 CI 3.30 5.98 2.76 3.17 5.98 2.70 3.11 5.98 2.59

TABLE 19.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3380 4600 3530 4530 3530 4530 Average 3630 5170 3680 5130 3690 5090 Stdev 2270 3180 2260 3190 2260 3190 p (t-test) 0.0028 0.0052 0.0075 Min 62.8 413 62.8 413 62.8 413 Max 10700 21200 10700 21200 10700 21200 n (Patient) 49 102 48 103 46 105 sCr only Median 3700 4680 3700 4680 3730 4670 Average 3750 5170 3750 5170 3800 5120 Stdev 2150 3280 2150 3280 2140 3290 p (t-test) 0.0056 0.0056 0.010 Min 276 62.8 276 62.8 276 62.8 Max 10700 21200 10700 21200 10700 21200 n (Patient) 52 98 52 98 51 99 UO only Median 4150 4470 4120 4470 4050 4670 Average 4550 5000 4600 4860 4430 5100 Stdev 2740 3510 2760 3390 2640 3460 p (t-test) 0.40 0.61 0.18 Min 62.8 733 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 104 46 97 53 91 59 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.63 0.52 0.64 0.63 0.51 0.64 0.62 0.55 SE 0.046 0.046 0.052 0.047 0.046 0.050 0.047 0.047 0.048 p Value 0.0010 0.0045 0.67 0.0027 0.0045 0.80 0.0037 0.011 0.30 nCohort 49 52 104 48 52 97 46 51 91 Recovered nCohort 102 98 46 103 98 53 105 99 59 Non-recovered Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 79% 78% 76% 79% 78% 75% 78% 77% 76% Specificity 35% 31% 26% 33% 31% 26% 33% 29% 26% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 58% 57% 52% 57% 57% 53% 57% 57% 56% Specificity 65% 63% 51% 65% 63% 52% 65% 63% 54% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 31% 33% 24% 31% 33% 23% 30% 32% 25% Specificity 88% 88% 74% 88% 88% 73% 87% 88% 75% OR Quartile 2 2.05 1.54 1.12 1.84 1.54 1.07 1.73 1.38 1.15 p Value 0.064 0.27 0.79 0.12 0.27 0.87 0.17 0.41 0.72 Lower limit 0.959 0.721 0.498 0.858 0.721 0.493 0.798 0.643 0.539 of 95% CI Upper limit 4.38 3.27 2.50 3.95 3.27 2.32 3.73 2.95 2.46 of 95% CI OR Quartile 3 2.58 2.32 1.13 2.45 2.32 1.19 2.50 2.19 1.48 p Value 0.0086 0.017 0.72 0.013 0.017 0.61 0.013 0.026 0.24 Lower limit 1.27 1.16 0.566 1.20 1.16 0.609 1.22 1.10 0.766 of 95% CI Upper limit 5.24 4.63 2.27 4.97 4.63 2.33 5.13 4.39 2.86 of 95% CI OR Quartile 4 3.28 3.72 0.896 3.15 3.72 0.799 2.92 3.58 1.01 p Value 0.014 0.0067 0.79 0.018 0.0067 0.58 0.027 0.0085 0.98 Lower limit 1.27 1.44 0.400 1.22 1.44 0.365 1.13 1.38 0.475 of 95% CI Upper limit 8.48 9.61 2.01 8.17 9.61 1.75 7.58 9.26 2.14 of 95% CI

TABLE 19.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3860 4500 3860 4500 3860 4500 Average 4290 4950 4290 4950 4310 4910 Stdev 2640 3220 2640 3220 2640 3210 p (t-test) 0.18 0.18 0.23 Min 62.8 276 62.8 276 62.8 276 Max 11800 21200 11800 21200 11800 21200 n (Patient) 64 87 64 87 62 89 sCr only Median 3940 4490 3940 4490 4000 4480 Average 4380 4890 4380 4890 4430 4850 Stdev 2530 3320 2530 3320 2520 3330 p (t-test) 0.30 0.30 0.40 Min 703 62.8 703 62.8 703 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 67 84 67 84 66 85 UO only Median 4060 4710 4050 4710 3930 4780 Average 4550 5020 4540 5010 4420 5170 Stdev 2710 3580 2730 3500 2620 3520 p (t-test) 0.37 0.37 0.14 Min 62.8 276 62.8 276 62.8 276 Max 13800 21200 13800 21200 13800 21200 n (Patient) 105 45 103 47 95 55 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.56 0.54 0.54 0.56 0.54 0.54 0.56 0.53 0.56 SE 0.047 0.047 0.052 0.047 0.047 0.051 0.047 0.047 0.049 p Value 0.18 0.41 0.50 0.18 0.41 0.43 0.22 0.57 0.19 nCohort 64 67 105 64 67 103 62 66 95 Recovered nCohort 87 84 45 87 84 47 89 85 55 Non-recovered Cutoff 2740 2740 2740 2740 2740 2740 2740 2740 2740 Quartile 2 Sensitivity 76% 74% 73% 76% 74% 74% 75% 73% 75% Specificity 27% 24% 25% 27% 24% 25% 26% 23% 25% Cutoff 4230 4230 4260 4230 4230 4260 4230 4230 4260 Quartile 3 Sensitivity 57% 56% 58% 57% 56% 60% 57% 55% 60% Specificity 59% 57% 53% 59% 57% 54% 60% 56% 56% Cutoff 6010 6010 6060 6010 6010 6060 6010 6010 6060 Quartile 4 Sensitivity 28% 29% 27% 28% 29% 26% 27% 28% 27% Specificity 78% 79% 75% 78% 79% 75% 77% 79% 76% OR Quartile 2 1.14 0.884 0.905 1.14 0.884 0.985 1.06 0.793 0.990 p Value 0.73 0.75 0.81 0.73 0.75 0.97 0.88 0.54 0.98 Lower limit 0.542 0.421 0.408 0.542 0.421 0.446 0.503 0.375 0.462 of 95% CI Upper limit 2.38 1.86 2.01 2.38 1.86 2.17 2.23 1.68 2.12 of 95% CI OR Quartile 3 1.98 1.66 1.56 1.98 1.66 1.76 1.99 1.58 1.89 p Value 0.042 0.12 0.21 0.042 0.12 0.11 0.041 0.17 0.064 Lower limit 1.03 0.871 0.773 1.03 0.871 0.872 1.03 0.826 0.964 of 95% CI Upper limit 3.80 3.18 3.16 3.80 3.18 3.54 3.84 3.01 3.72 of 95% CI OR Quartile 4 1.36 1.51 1.10 1.36 1.51 1.02 1.27 1.46 1.17 p Value 0.43 0.28 0.81 0.43 0.28 0.97 0.54 0.33 0.68 Lower limit 0.639 0.711 0.499 0.639 0.711 0.460 0.594 0.686 0.551 of 95% CI Upper limit 2.90 3.22 2.45 2.90 3.22 2.24 2.70 3.11 2.50 of 95% CI

TABLE 19.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered'’ and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3790 4710 3860 4700 3770 4700 Average 4210 5090 4250 5040 4230 5040 Stdev 2560 3310 2550 3320 2580 3280 p (t-test) 0.072 0.11 0.095 Min 62.8 276 62.8 276 62.8 276 Max 11800 21200 11800 21200 11800 21200 n (Patient) 73 78 72 79 70 81 sCr only Median 3930 4670 3930 4670 3930 4670 Average 4300 5020 4300 5020 4300 5020 Stdev 2500 3380 2500 3380 2500 3380 p (t-test) 0.14 0.14 0.14 Min 703 62.8 703 62.8 703 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 74 77 74 77 74 77 UO only Median 4050 4800 4050 4800 3930 4820 Average 4460 5230 4470 5170 4390 5230 Stdev 2610 3710 2630 3650 2580 3570 p (t-test) 0.15 0.19 0.096 Min 62.8 276 62.8 276 62.8 276 Max 13800 21200 13800 21200 13800 21200 n (Patient) 105 45 103 47 96 54 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.56 0.56 0.57 0.56 0.55 0.58 0.56 0.58 SE 0.046 0.047 0.052 0.046 0.047 0.051 0.046 0.047 0.049 p Value 0.073 0.20 0.25 0.12 0.20 0.28 0.086 0.20 0.13 nCohort 73 74 105 72 74 103 70 74 96 Recovered nCohort 78 77 45 79 77 47 81 77 54 Non-recovered Cutoff 2740 2740 2740 2740 2740 2740 2740 2740 2740 Quartile 2 Sensitivity 77% 75% 73% 76% 75% 72% 77% 75% 74% Specificity 27% 26% 25% 26% 26% 24% 27% 26% 25% Cutoff 4230 4230 4260 4230 4230 4260 4230 4230 4260 Quartile 3 Sensitivity 60% 58% 62% 59% 58% 62% 60% 58% 63% Specificity 60% 58% 55% 60% 58% 55% 61% 58% 57% Cutoff 6010 6010 6060 6010 6010 6060 6010 6010 6060 Quartile 4 Sensitivity 29% 30% 29% 29% 30% 28% 28% 30% 28% Specificity 79% 80% 76% 79% 80% 76% 79% 80% 76% OR Quartile 2 1.26 1.05 0.905 1.13 1.05 0.838 1.22 1.05 0.952 p Value 0.54 0.89 0.81 0.74 0.89 0.66 0.60 0.89 0.90 Lower limit 0.602 0.506 0.408 0.543 0.506 0.384 0.582 0.506 0.444 of 95% CI Upper limit 2.63 2.20 2.01 2.36 2.20 1.83 2.54 2.20 2.04 of 95% CI OR Quartile 3 2.30 1.95 2.03 2.18 1.95 2.00 2.44 1.95 2.28 p Value 0.012 0.043 0.052 0.019 0.043 0.055 0.0077 0.043 0.018 Lower limit 1.20 1.02 0.994 1.14 1.02 0.987 1.27 1.02 1.15 of 95% CI Upper limit 4.42 3.73 4.15 4.18 3.73 4.04 4.70 3.73 4.52 of 95% CI OR Quartile 4 1.62 1.68 1.30 1.56 1.68 1.19 1.45 1.68 1.22 p Value 0.21 0.18 0.51 0.24 0.18 0.66 0.33 0.18 0.61 Lower limit 0.765 0.793 0.593 0.739 0.793 0.546 0.688 0.793 0.572 of 95% CI Upper limit 3.42 3.54 2.85 3.30 3.54 2.61 3.07 3.54 2.60 of 95% CI

TABLE 19.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3860 4710 3920 4690 3790 4690 Average 4250 5090 4290 5040 4260 5040 Stdev 2560 3350 2550 3360 2580 3310 p (t-test) 0.083 0.12 0.11 Min 62.8 276 62.8 276 62.8 276 Max 11800 21200 11800 21200 11800 21200 n (Patient) 76 75 75 76 73 78 sCr only Median 3930 4580 3930 4580 3930 4580 Average 4320 5030 4320 5030 4320 5030 Stdev 2500 3420 2500 3420 2500 3420 p (t-test) 0.15 0.15 0.15 Min 703 62.8 703 62.8 703 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 77 74 77 74 77 74 UO only Median 3930 4820 3930 4820 3920 4840 Average 4460 5160 4460 5160 4380 5230 Stdev 2670 3540 2670 3540 2600 3530 p (t-test) 0.18 0.18 0.092 Min 62.8 276 62.8 276 62.8 276 Max 13800 21200 13800 21200 13800 21200 n (Patient) 100 50 100 50 95 55 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.56 0.56 0.57 0.56 0.56 0.57 0.56 0.58 SE 0.046 0.047 0.050 0.047 0.047 0.050 0.046 0.047 0.049 p Value 0.093 0.22 0.20 0.15 0.22 0.20 0.11 0.22 0.10 nCohort 76 77 100 75 77 100 73 77 95 Recovered nCohort 75 74 50 76 74 50 78 74 55 Non-recovered Cutoff 2740 2740 2740 2740 2740 2740 2740 2740 2740 Quartile 2 Sensitivity 77% 76% 74% 76% 76% 74% 77% 76% 75% Specificity 28% 26% 25% 27% 26% 25% 27% 26% 25% Cutoff 4230 4230 4260 4230 4230 4260 4230 4230 4260 Quartile 3 Sensitivity 60% 58% 62% 59% 58% 62% 60% 58% 64% Specificity 59% 57% 56% 59% 57% 56% 60% 57% 58% Cutoff 6010 6010 6060 6010 6010 6060 6010 6010 6060 Quartile 4 Sensitivity 29% 30% 26% 29% 30% 26% 28% 30% 27% Specificity 79% 79% 75% 79% 79% 75% 78% 79% 76% OR Quartile 2 1.30 1.09 0.949 1.17 1.09 0.949 1.26 1.09 0.990 p Value 0.48 0.82 0.89 0.67 0.82 0.89 0.54 0.82 0.98 Lower limit 0.623 0.523 0.436 0.561 0.523 0.436 0.602 0.523 0.462 of 95% CI Upper limit 2.73 2.28 2.06 2.45 2.28 2.06 2.63 2.28 2.12 of 95% CI OR Quartile 3 2.18 1.85 2.08 2.06 1.85 2.08 2.30 1.85 2.41 p Value 0.019 0.062 0.039 0.029 0.062 0.039 0.012 0.062 0.012 Lower limit 1.14 0.970 1.04 1.08 0.970 1.04 1.20 0.970 1.21 of 95% CI Upper limit 4.17 3.53 4.16 3.94 3.53 4.16 4.42 3.53 4.77 of 95% CI OR Quartile 4 1.56 1.61 1.05 1.50 1.61 1.05 1.40 1.61 1.17 p Value 0.24 0.21 0.89 0.28 0.21 0.89 0.37 0.21 0.68 Lower limit 0.741 0.768 0.484 0.715 0.768 0.484 0.666 0.768 0.551 of 95% CI Upper limit 3.27 3.39 2.29 3.16 3.39 2.29 2.94 3.39 2.50 of 95% CI

TABLE 19.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 4060 4580 4090 4500 4060 4490 Average 4540 4840 4580 4780 4580 4780 Stdev 2720 3370 2710 3380 2730 3350 p (t-test) 0.55 0.68 0.69 Min 703 62.8 703 62.8 703 62.8 Max 13800 21200 13800 21200 13800 21200 n (Patient) 89 62 88 63 87 64 sCr only Median 3930 4690 3930 4690 3930 4690 Average 4310 5060 4310 5060 4310 5060 Stdev 2520 3420 2520 3420 2520 3420 p (t-test) 0.12 0.12 0.12 Min 703 62.8 703 62.8 703 62.8 Max 11800 21200 11800 21200 11800 21200 n (Patient) 79 72 79 72 79 72 UO only Median 3930 4790 3930 4790 3930 4750 Average 4480 5080 4480 5080 4520 5000 Stdev 2670 3520 2670 3520 2680 3490 p (t-test) 0.24 0.24 0.34 Min 413 62.8 413 62.8 413 62.8 Max 13800 21200 13800 21200 13800 21200 n (Patient) 98 52 98 52 96 54 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.52 0.56 0.56 0.51 0.56 0.56 0.51 0.56 0.54 SE 0.048 0.047 0.050 0.048 0.047 0.050 0.048 0.047 0.049 p Value 0.61 0.17 0.27 0.78 0.17 0.27 0.77 0.17 0.38 nCohort 89 79 98 88 79 98 87 79 96 Recovered nCohort 62 72 52 63 72 52 64 72 54 Non-recovered Cutoff 2740 2740 2740 2740 2740 2740 2740 2740 2740 Quartile 2 Sensitivity 74% 78% 75% 73% 78% 75% 73% 78% 74% Specificity 25% 28% 26% 24% 28% 26% 24% 28% 25% Cutoff 4230 4230 4260 4230 4230 4260 4230 4230 4260 Quartile 3 Sensitivity 56% 58% 60% 56% 58% 60% 56% 58% 59% Specificity 54% 57% 55% 53% 57% 55% 54% 57% 55% Cutoff 6010 6010 6060 6010 6010 6060 6010 6010 6060 Quartile 4 Sensitivity 24% 28% 25% 24% 28% 25% 23% 28% 24% Specificity 74% 77% 74% 74% 77% 74% 74% 77% 74% OR Quartile 2 0.944 1.35 1.03 0.848 1.35 1.03 0.880 1.35 0.952 p Value 0.88 0.43 0.95 0.66 0.43 0.95 0.73 0.43 0.90 Lower limit 0.448 0.643 0.473 0.404 0.643 0.473 0.419 0.643 0.444 of 95% CI Upper limit 1.99 2.84 2.23 1.78 2.84 2.23 1.85 2.84 2.04 of 95% CI OR Quartile 3 1.52 1.85 1.81 1.43 1.85 1.81 1.51 1.85 1.79 p Value 0.21 0.061 0.088 0.28 0.061 0.088 0.21 0.061 0.090 Lower limit 0.790 0.971 0.916 0.748 0.971 0.916 0.789 0.971 0.912 of 95% CI Upper limit 2.91 3.54 3.58 2.74 3.54 3.58 2.89 3.54 3.52 of 95% CI OR Quartile 4 0.916 1.30 0.973 0.883 1.30 0.973 0.852 1.30 0.900 p Value 0.82 0.48 0.95 0.75 0.48 0.95 0.67 0.48 0.79 Lower limit 0.432 0.624 0.449 0.417 0.624 0.449 0.403 0.624 0.416 of 95% CI Upper limit 1.94 2.72 2.11 1.87 2.72 2.11 1.80 2.72 1.95 of 95% CI

Example 20. Use of C—C Motif Chemokine 15 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 15 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 20.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 65.6 585 65.6 770 77.7 784 Average 1180 2680 1140 3040 1460 2460 Stdev 5100 6030 4920 6440 5910 3940 p (t-test) 0.020 0.0044 0.16 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 19200 n (Patient) 204 112 220 96 235 81 sCr only Median 65.2 604 65.6 803 75.3 791 Average 1170 2770 1110 3170 1450 2490 Stdev 5060 6130 4890 6520 5920 3950 p (t-test) 0.014 0.0023 0.14 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 19200 n (Patient) 207 108 222 93 234 81 UO only Median 130 344 130 406 129 491 Average 1750 1550 1730 1670 1720 1720 Stdev 5760 3230 5730 3330 5720 3370 p (t-test) 0.83 0.95 1.00 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 18400 53500 18400 53500 18400 n (Patient) 274 40 277 37 278 36 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.71 0.58 0.73 0.74 0.60 0.72 0.72 0.61 SE 0.032 0.032 0.050 0.033 0.033 0.052 0.035 0.035 0.052 p Value 3.6E−10 2.6E−11 0.12 4.4E−12 2.8E−13 0.065 1.0E−9 6.9E−10 0.034 nCohort 204 207 274 220 222 277 235 234 278 Non-persistent nCohort 112 108 40 96 93 37 81 81 36 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 88% 90% 82% 90% 91% 84% 90% 93% 86% Specificity 29% 30% 24% 29% 30% 24% 28% 29% 24% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 71% 71% 68% 75% 75% 70% 77% 74% 72% Specificity 61% 61% 53% 61% 60% 53% 59% 58% 53% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 42% 44% 32% 46% 48% 35% 46% 47% 36% Specificity 84% 85% 76% 84% 85% 76% 82% 82% 76% OR Quartile 2 2.92 3.86 1.50 3.53 4.50 1.65 3.56 5.12 2.01 p Value 9.7E−4 1.3E−4 0.36 5.6E−4 1.6E−4 0.29 0.0015 2.7E−4 0.16 Lower limit 1.54 1.93 0.632 1.72 2.06 0.659 1.63 2.13 0.751 of 95% CI Upper limit 5.51 7.69 3.54 7.22 9.81 4.12 7.80 12.3 5.37 of 95% CI OR Quartile 3 3.79 3.86 2.30 4.67 4.63 2.63 4.72 3.97 2.92 p Value 1.3E−7 1.3E−7 0.020 1.7E−8 3.0E−8 0.011 1.3E−7 1.5E−6 0.0062 Lower limit 2.31 2.34 1.14 2.74 2.69 1.25 2.66 2.26 1.36 of 95% CI Upper limit 6.21 6.38 4.65 7.99 7.97 5.54 8.41 6.95 6.28 of 95% CI OR Quartile 4 3.89 4.21 1.52 4.47 5.18 1.73 3.86 4.16 1.82 p Value 5.7E−7 1.4E−7 0.25 5.4E−8 3.6E−9 0.14 1.5E−6 4.0E−7 0.11 Lower limit 2.28 2.47 0.741 2.61 3.00 0.835 2.23 2.40 0.871 of 95% CI Upper limit 6.62 7.20 3.11 7.68 8.95 3.59 6.70 7.22 3.78 of 95% CI

TABLE 20.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 59.2 576 59.2 618 65.6 616 Average 1170 2490 1100 2820 1460 2290 Stdev 5270 5700 5040 6070 6080 3820 p (t-test) 0.034 0.0075 0.21 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 19200 n (Patient) 185 131 203 113 218 98 sCr only Median 60.0 577 59.2 717 65.2 616 Average 1140 2610 1090 2900 1460 2300 Stdev 5190 5840 5020 6140 6090 3820 p (t-test) 0.021 0.0050 0.21 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 19200 n (Patient) 191 124 205 110 217 98 UO only Median 118 406 118 488 118 521 Average 1790 1450 1760 1530 1760 1560 Stdev 5950 2870 5910 2960 5900 2980 p (t-test) 0.67 0.78 0.81 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 18400 53500 18400 53500 18400 n (Patient) 255 59 259 55 260 54 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.71 0.60 0.73 0.73 0.61 0.71 0.71 0.62 SE 0.030 0.031 0.042 0.031 0.031 0.044 0.033 0.033 0.044 p Value 2.7E−11 1.4E−11 0.020 2.3E−13 3.9E−14 0.014 3.3E−10 2.8E−10 0.0069 nCohort 185 191 255 203 205 259 218 217 260 Non-persistent nCohort 131 124 59 113 110 55 98 98 54 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 87% 89% 85% 89% 91% 85% 89% 91% 87% Specificity 31% 31% 25% 31% 31% 25% 29% 30% 25% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 70% 70% 68% 73% 74% 69% 74% 72% 70% Specificity 64% 63% 54% 63% 62% 54% 61% 60% 54% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 40% 42% 31% 43% 45% 33% 42% 43% 33% Specificity 85% 86% 76% 85% 86% 76% 83% 83% 77% OR Quartile 2 2.99 3.60 1.86 3.70 4.54 1.97 3.21 4.23 2.28 p Value 3.3E−4 7.6E−5 0.11 1.3E−4 3.3E−5 0.097 9.4E−4 1.5E−4 0.054 Lower limit 1.64 1.91 0.867 1.90 2.22 0.884 1.61 2.01 0.984 of 95% CI Upper limit 5.43 6.79 4.00 7.22 9.27 4.38 6.42 8.90 5.30 of 95% CI OR Quartile 3 4.25 3.97 2.48 4.72 4.64 2.63 4.57 3.93 2.81 p Value 3.5E−9 2.3E−8 0.0029 1.8E−9 3.5E−9 0.0023 1.9E−8 2.5E−7 0.0014 Lower limit 2.63 2.45 1.36 2.85 2.79 1.41 2.69 2.34 1.49 of 95% CI Upper limit 6.88 6.45 4.52 7.83 7.73 4.90 7.76 6.61 5.30 of 95% CI OR Quartile 4 3.85 4.39 1.40 4.42 5.06 1.58 3.41 3.65 1.63 p Value 8.8E−7 8.6E−8 0.29 6.1E−8 5.0E−9 0.16 6.4E−6 2.0E−6 0.13 Lower limit 2.25 2.55 0.748 2.58 2.94 0.839 2.00 2.14 0.865 of 95% CI Upper limit 6.59 7.54 2.61 7.56 8.71 2.97 5.80 6.22 3.08 of 95% CI

TABLE 20.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 55.7 585 55.3 692 63.1 635 Average 1100 2470 1030 2740 1400 2290 Stdev 5340 5580 5130 5840 6200 3820 p (t-test) 0.026 0.0065 0.17 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 19200 n (Patient) 174 142 189 127 204 112 sCr only Median 56.1 596 55.7 784 65.0 616 Average 1070 2600 1020 2850 1390 2350 Stdev 5240 5720 5070 5960 6150 3880 p (t-test) 0.014 0.0038 0.14 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 19200 n (Patient) 181 134 194 121 207 108 UO only Median 118 406 118 521 118 521 Average 1780 1510 1750 1610 1750 1610 Stdev 6040 2930 5980 3010 5980 3010 p (t-test) 0.72 0.86 0.86 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 18400 53500 18400 53500 18400 n (Patient) 245 69 250 64 250 64 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.72 0.59 0.74 0.74 0.61 0.71 0.72 0.61 SE 0.029 0.029 0.040 0.029 0.030 0.041 0.031 0.032 0.041 p Value 3.3E−13 4.0E−14 0.018 4.4E−16 2.2E−16 0.0070 1.2E−11 1.1E−11 0.0070 nCohort 174 181 245 189 194 250 204 207 250 Non-persistent nCohort 142 134 69 127 121 64 112 108 64 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 87% 89% 84% 89% 91% 86% 88% 91% 86% Specificity 32% 33% 25% 32% 32% 26% 30% 31% 26% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 70% 70% 65% 72% 73% 67% 72% 71% 67% Specificity 66% 65% 54% 65% 64% 54% 62% 61% 54% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 40% 43% 30% 44% 46% 33% 42% 44% 33% Specificity 87% 88% 76% 88% 88% 77% 84% 85% 77% OR Quartile 2 2.99 3.84 1.79 3.75 4.81 2.10 3.25 4.39 2.10 p Value 2.1E−4 2.2E−5 0.11 4.3E−5 7.9E−6 0.055 3.9E−4 5.0E−5 0.055 Lower limit 1.68 2.06 0.882 1.99 2.42 0.984 1.69 2.15 0.984 of 95% CI Upper limit 5.34 7.14 3.62 7.08 9.58 4.50 6.23 8.96 4.50 of 95% CI OR Quartile 3 4.49 4.30 2.23 4.90 4.72 2.44 4.31 3.86 2.44 p Value 6.4E−10 2.5E−9 0.0047 2.2E−10 8.4E−10 0.0025 1.1E−8 1.3E−7 0.0025 Lower limit 2.79 2.66 1.28 3.00 2.88 1.37 2.61 2.34 1.37 of 95% CI Upper limit 7.22 6.94 3.88 8.00 7.76 4.35 7.12 6.38 4.35 of 95% CI OR Quartile 4 4.63 5.35 1.41 5.69 6.41 1.62 3.89 4.21 1.62 p Value 7.6E−8 5.0E−9 0.25 1.31E−9 1.0E−10 0.12 5.7E−7 1.4E−7 0.12 Lower limit 2.65 3.05 0.781 3.25 3.65 0.888 2.28 2.47 0.888 of 95% CI Upper limit 8.10 9.39 2.55 9.96 11.2 2.94 6.62 7.20 2.94 of 95% CI

TABLE 20.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent'’ and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 54.2 585 54.2 671 60.6 616 Average 1100 2450 1040 2720 1160 2670 Stdev 5380 5540 5150 5820 5100 6000 p (t-test) 0.029 0.0073 0.019 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 52500 n (Patient) 172 144 188 128 200 116 sCr only Median 56.1 585 55.3 701 61.1 616 Average 1080 2560 1030 2790 1150 2740 Stdev 5270 5680 5110 5900 5060 6090 p (t-test) 0.017 0.0053 0.014 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 52500 53500 52500 53500 52500 n (Patient) 179 136 191 124 203 112 UO only Median 116 351 116 380 116 380 Average 1750 1650 1710 1760 1710 1760 Stdev 6010 3470 5940 3590 5940 3590 p (t-test) 0.90 0.95 0.95 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 238 76 244 70 244 70 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.72 0.60 0.74 0.74 0.61 0.71 0.72 0.61 SE 0.029 0.030 0.038 0.029 0.030 0.040 0.031 0.031 0.040 p Value 1.3E−13 2.7E−13 0.011 6.7E−16 6.7E−16 0.0061 7.6E−12 6.2E−12 0.0061 nCohort 172 179 238 188 191 244 200 203 244 Non-persistent nCohort 144 136 76 128 124 70 116 112 70 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 87% 88% 84% 89% 90% 86% 89% 90% 86% Specificity 32% 32% 26% 32% 32% 26% 30% 31% 26% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 69% 69% 66% 72% 72% 67% 71% 71% 67% Specificity 66% 64% 55% 65% 64% 55% 62% 61% 55% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 40% 42% 30% 44% 45% 31% 41% 43% 31% Specificity 88% 88% 76% 88% 88% 77% 84% 85% 77% OR Quartile 2 3.09 3.60 1.84 3.82 4.49 2.09 3.48 4.13 2.09 p Value 1.3E−4 3.7E−5 0.080 3.5E−5 1.1E−5 0.047 1.7E−4 5.5E−5 0.047 Lower limit 1.73 1.96 0.930 2.02 2.30 1.01 1.81 2.07 1.01 of 95% CI Upper limit 5.52 6.61 3.63 7.20 8.75 4.33 6.67 8.24 4.33 of 95% CI OR Quartile 3 4.47 4.02 2.35 4.72 4.50 2.49 3.93 3.76 2.49 p Value 6.6E−10 9.4E−9 0.0018 4.5E−10 1.8E−9 0.0014 4.6E−8 1.6E−7 0.0014 Lower limit 2.78 2.50 1.37 2.90 2.75 1.42 2.41 2.29 1.42 of 95% CI Upper limit 7.18 6.47 4.03 7.70 7.34 4.35 6.43 6.16 4.35 of 95% CI OR Quartile 4 4.85 5.15 1.41 5.58 6.02 1.50 3.85 4.16 1.50 p Value 4.3E−8 1.1E−8 0.24 1.6E−9 3.7E−10 0.17 6.9E−7 1.8E−7 0.17 Lower limit 2.76 2.94 0.795 3.19 3.43 0.837 2.26 2.44 0.837 of 95% CI Upper limit 8.53 9.03 2.50 9.76 10.5 2.70 6.55 7.11 1 2.70 of 95% CI

TABLE 20.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 54.2 585 53.2 692 59.6 616 Average 798 2760 754 3040 1170 2600 Stdev 3610 6900 3460 7220 5150 5910 p (t-test) 0.0014 2.2E−4 0.024 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 53500 52500 n (Patient) 168 148 183 133 196 120 sCr only Median 56.1 585 55.3 701 61.1 616 Average 787 2890 752 3130 899 3130 Stdev 3540 7070 3430 7350 3490 7630 p (t-test) 6.9E−4 1.4E−4 4.7E−4 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 175 140 187 128 199 116 UO only Median 106 406 104 447 104 447 Average 1730 1710 1710 1760 1710 1760 Stdev 6070 3420 6030 3470 6030 3470 p (t-test) 0.98 0.94 0.94 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 233 81 236 78 236 78 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.72 0.62 0.75 0.74 0.62 0.72 0.72 0.62 SE 0.029 0.029 0.037 0.029 0.029 0.038 0.031 0.031 0.038 p Value 3.4E−14 1.0E−13 0.0019 0 2.2E−16 0.0012 2.3E−12 3.5E−12 0.0012 nCohort 168 175 233 183 187 236 196 199 236 Non-persistent nCohort 148 140 81 133 128 78 120 116 78 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 86% 87% 85% 89% 89% 86% 89% 89% 86% Specificity 32% 32% 26% 33% 32% 26% 31% 31% 26% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 69% 69% 68% 72% 71% 69% 71% 70% 69% Specificity 67% 65% 56% 66% 64% 56% 63% 61% 56% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 41% 42% 32% 44% 45% 33% 42% 43% 33% Specificity 89% 89% 77% 89% 89% 78% 85% 85% 78% OR Quartile 2 3.03 3.19 2.04 4.15 3.85 2.17 3.72 3.50 2.17 p Value 1.4E−4 1.1E−4 0.040 1.1E−5 3.1E−5 0.030 7.5E−5 1.6E−4 0.030 Lower limit 1.71 1.77 1.03 2.20 2.04 1.08 1.94 1.83 1.08 of 95% CI Upper limit 5.37 5.74 4.02 7.82 7.25 4.37 7.13 6.71 4.37 of 95% CI OR Quartile 3 4.43 3.98 2.72 5.06 4.41 2.91 4.09 3.67 2.91 p Value 6.9E−10 1.0E−8 2.4E−4 6.9E−11 2.1E−9 1.3E−4 1.6E−8 1.8E−7 1.3E−4 Lower limit 2.76 2.48 1.59 3.11 2.71 1.68 2.51 2.25 1.68 of 95% CI Upper limit 7.12 6.38 4.63 8.24 7.16 5.01 6.67 5.98 5.01 of 95% CI OR Quartile 4 5.35 5.65 1.61 6.50 6.55 1.73 4.11 4.44 1.73 p Value 1.4E−8 3.4E−9 0.096 2.0E−10 1.2E−10 0.057 2.3E−7 5.8E−8 0.057 Lower limit 3.00 3.18 0.919 3.65 3.70 0.985 2.41 2.59 0.985 of 95% CI Upper limit 9.54 10.0 2.81 11.6 11.6 3.03 7.03 7.61 3.03 of 95% CI

TABLE 20.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 4090 4500 4090 4500 4170 4300 Average 4460 5020 4470 5090 4630 4780 Stdev 2710 3430 2690 3590 2790 3560 p (t-test) 0.27 0.24 0.78 Min 62.8 413 62.8 413 62.8 413 Max 11800 21200 11800 21200 13800 21200 n (Patient) 96 55 104 47 111 40 sCr only Median 4150 4490 4120 4500 4230 4300 Average 4520 4930 4480 5090 4660 4720 Stdev 2700 3470 2660 3650 2780 3560 p (t-test) 0.42 0.26 0.91 Min 276 62.8 276 62.8 276 62.8 Max 11800 21200 11800 21200 13800 21200 n (Patient) 94 56 103 47 108 42 UO only Median 4170 4800 4170 4800 4150 4880 Average 4560 5570 4560 5570 4540 5820 Stdev 2670 4660 2670 4660 2680 4660 p (t-test) 0.17 0.17 0.089 Min 62.8 1120 62.8 1120 62.8 1630 Max 13800 21200 13800 21200 13800 21200 n (Patient) 131 19 131 19 132 18 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.54 0.52 0.54 0.54 0.54 0.54 0.49 0.49 0.57 SE 0.049 0.049 0.072 0.051 0.051 0.072 0.054 0.053 0.074 p Value 0.44 0.65 0.54 0.43 0.43 0.54 0.89 0.80 0.32 nCohort 96 94 131 104 103 131 111 108 132 Non-persistent nCohort 55 56 19 47 47 19 40 42 18 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 75% 73% 68% 79% 79% 68% 78% 76% 72% Specificity 25% 24% 24% 27% 27% 24% 26% 26% 25% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 56% 54% 63% 57% 55% 63% 52% 50% 67% Specificity 53% 52% 52% 53% 52% 52% 50% 50% 52% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 29% 29% 26% 28% 30% 26% 20% 21% 28% Specificity 77% 77% 75% 76% 77% 75% 73% 73% 75% OR Quartile 2 0.976 0.885 0.700 1.36 1.38 0.700 1.22 1.12 0.867 p Value 0.95 0.75 0.50 0.46 0.44 0.50 0.65 0.79 0.80 Lower limit 0.455 0.416 0.246 0.599 0.607 0.246 0.518 0.488 0.287 of 95% CI Upper limit 2.09 1.89 1.99 3.10 3.14 1.99 2.86 2.57 2.61 of 95% CI OR Quartile 3 1.46 1.26 1.85 1.52 1.36 1.85 1.13 1.00 2.19 p Value 0.26 0.50 0.22 0.24 0.38 0.22 0.75 1.0 0.14 Lower limit 0.751 0.647 0.685 0.756 0.682 0.685 0.546 0.490 0.776 of 95% CI Upper limit 2.85 2.44 5.00 3.04 2.73 5.00 2.32 2.04 6.18 of 95% CI OR Quartile 4 1.38 1.31 1.06 1.21 1.40 1.06 0.675 0.743 1.15 p Value 0.40 0.48 0.92 0.64 0.40 0.92 0.38 0.49 0.80 Lower limit 0.651 0.618 0.355 0.553 0.644 0.355 0.280 0.317 0.383 of 95% CI Upper limit 2.93 2.77 3.17 2.64 3.03 3.17 1.63 1.74 3.48 of 95% CI

TABLE 20.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 4000 4480 4000 4480 4170 4420 Average 4440 4980 4480 4980 4640 4720 Stdev 2720 3350 2750 3360 2850 3300 p (t-test) 0.28 0.32 0.88 Min 62.8 413 62.8 413 62.8 413 Max 11800 21200 11800 21200 13800 21200 n (Patient) 88 63 94 57 101 50 sCr only Median 4090 4490 4060 4500 4170 4370 Average 4490 4930 4460 5020 4640 4730 Stdev 2680 3430 2650 3500 2770 3440 p (t-test) 0.38 0.26 0.87 Min 276 62.8 276 62.8 276 62.8 Max 11800 21200 11800 21200 13800 21200 n (Patient) 88 62 93 57 99 51 UO only Median 4170 4670 4170 4670 4150 4690 Average 4600 5110 4600 5110 4570 5260 Stdev 2710 4090 2710 4090 2710 4090 p (t-test) 0.42 0.42 0.29 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 123 27 123 27 124 26 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.54 0.53 0.52 0.54 0.54 0.52 0.50 0.49 0.54 SE 0.048 0.048 0.062 0.049 0.049 0.062 0.050 0.050 0.063 p Value 0.39 0.55 0.80 0.39 0.39 0.80 0.98 0.89 0.57 nCohort 88 88 123 94 93 123 101 99 124 Non-persistent nCohort 63 62 27 57 57 27 50 51 26 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 76% 74% 70% 79% 77% 70% 78% 75% 73% Specificity 26% 25% 24% 28% 27% 24% 27% 25% 25% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 57% 55% 59% 58% 56% 59% 54% 51% 62% Specificity 55% 53% 52% 54% 54% 52% 51% 51% 52% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 27% 27% 22% 26% 28% 22% 20% 22% 23% Specificity 76% 76% 74% 76% 76% 74% 72% 73% 74% OR Quartile 2 1.13 0.958 0.766 1.43 1.24 0.766 1.29 0.988 0.905 p Value 0.75 0.91 0.57 0.37 0.58 0.57 0.53 0.97 0.84 Lower limit 0.535 0.455 0.304 0.657 0.576 0.304 0.581 0.455 0.347 of 95% CI Upper limit 2.40 2.02 1.93 3.13 2.69 1.93 2.88 2.15 2.36 of 95% CI OR Quartile 3 1.60 1.39 1.58 1.63 1.49 1.58 1.25 1.06 1.76 p Value 0.16 0.32 0.29 0.15 0.24 0.29 0.53 0.86 0.20 Lower limit 0.834 0.725 0.678 0.839 0.767 0.678 0.632 0.540 0.742 of 95% CI Upper limit 3.07 2.67 3.67 3.17 2.89 3.67 2.46 2.09 4.19 of 95% CI OR Quartile 4 1.18 1.21 0.812 1.10 1.26 0.812 0.652 0.733 0.862 p Value 0.66 0.62 0.68 0.80 0.55 0.68 0.31 0.45 0.77 Lower limit 0.562 0.574 0.301 0.519 0.595 0.301 0.287 0.329 0.318 of 95% CI Upper limit 2.47 2.53 2.19 2.34 2.67 2.19 1.48 1.63 2.34 of 95% CI

TABLE 20.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3930 4580 3930 4500 4090 4480 Average 4370 5030 4390 5030 4530 4880 Stdev 2750 3260 2730 3310 2830 3270 p (t-test) 0.18 0.20 0.49 Min 62.8 413 62.8 413 62.8 413 Max 11800 21200 11800 21200 13800 21200 n (Patient) 83 68 86 65 94 57 sCr only Median 3930 4580 3930 4580 4120 4480 Average 4420 4980 4430 5000 4590 4810 Stdev 2720 3310 2690 3380 2800 3330 p (t-test) 0.26 0.25 0.66 Min 276 62.8 276 62.8 276 62.8 Max 11800 21200 11800 21200 13800 21200 n (Patient) 82 68 86 64 93 57 UO only Median 4060 4780 4060 4780 4050 4790 Average 4550 5150 4550 5150 4520 5270 Stdev 2750 3690 2750 3690 2750 3680 p (t-test) 0.30 0.30 0.20 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 115 35 115 35 116 34 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.56 0.55 0.55 0.56 0.55 0.55 0.53 0.52 0.57 SE 0.047 0.047 0.057 0.047 0.048 0.057 0.049 0.049 0.057 p Value 0.18 0.27 0.37 0.22 0.29 0.37 0.51 0.73 0.23 nCohort 83 82 115 86 86 115 94 93 116 Non-persistent nCohort 68 68 35 65 64 35 57 57 34 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 78% 76% 74% 78% 77% 74% 79% 75% 76% Specificity 28% 27% 25% 28% 27% 25% 28% 26% 26% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 59% 57% 63% 58% 58% 63% 56% 54% 65% Specificity 57% 56% 54% 56% 56% 54% 53% 53% 54% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 28% 28% 26% 28% 28% 26% 25% 25% 26% Specificity 77% 77% 75% 77% 77% 75% 74% 74% 75% OR Quartile 2 1.35 1.19 0.974 1.41 1.19 0.974 1.43 1.07 1.13 p Value 0.43 0.64 0.95 0.37 0.65 0.95 0.37 0.86 0.78 Lower limit 0.641 0.567 0.409 0.662 0.563 0.409 0.657 0.499 0.463 of 95% CI Upper limit 2.86 2.51 2.32 3.00 2.52 2.32 3.13 2.29 2.77 of 95% CI OR Quartile 3 1.87 1.72 1.98 1.78 1.73 1.98 1.45 1.33 2.18 p Value 0.060 0.10 0.085 0.083 0.100 0.085 0.27 0.40 0.054 Lower limit 0.974 0.898 0.910 0.927 0.900 0.910 0.751 0.685 0.986 of 95% CI Upper limit 3.57 3.29 4.31 3.41 3.33 4.31 2.82 2.57 4.81 of 95% CI OR Quartile 4 1.31 1.29 1.03 1.26 1.29 1.03 0.950 0.936 1.08 p Value 0.48 0.50 0.95 0.53 0.50 0.95 0.89 0.86 0.86 Lower limit 0.625 0.615 0.431 0.604 0.616 0.431 0.444 0.437 0.452 of 95% CI Upper limit 2.73 2.69 2.44 2.65 2.71 2.44 2.03 2.00 2.58 of 95% CI

TABLE 20.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3920 4690 3930 4580 4060 4490 Average 4350 5040 4380 5040 4520 4880 Stdev 2780 3210 2740 3280 2860 3200 p (t-test) 0.16 0.18 0.48 Min 62.8 413 62.8 413 62.8 413 Max 11800 21200 11800 21200 13800 21200 n (Patient) 81 70 85 66 91 60 sCr only Median 3930 4670 3930 4580 4090 4490 Average 4410 4990 4430 4990 4570 4820 Stdev 2740 3280 2720 3330 2840 3260 p (t-test) 0.24 0.26 0.62 Min 276 62.8 276 62.8 276 62.8 Max 11800 21200 11800 21200 13800 21200 n (Patient) 81 69 84 66 90 60 UO only Median 3940 4800 3990 4790 3940 4800 Average 4500 5200 4510 5200 4480 5300 Stdev 2760 3510 2750 3560 2760 3540 p (t-test) 0.21 0.21 0.14 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 109 41 110 40 111 39 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.57 0.56 0.57 0.56 0.55 0.57 0.54 0.53 0.58 SE 0.047 0.047 0.054 0.047 0.048 0.054 0.048 0.048 0.054 p Value 0.12 0.22 0.20 0.18 0.28 0.23 0.44 0.60 0.14 nCohort 81 81 109 85 84 110 91 90 111 Non-persistent nCohort 70 69 41 66 66 40 60 60 39 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 79% 77% 76% 79% 77% 75% 78% 77% 77% Specificity 28% 27% 26% 28% 27% 25% 27% 27% 26% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 60% 58% 63% 59% 58% 62% 57% 55% 64% Specificity 58% 57% 55% 56% 56% 55% 54% 53% 55% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 27% 28% 24% 27% 27% 25% 25% 25% 26% Specificity 77% 77% 74% 76% 76% 75% 75% 74% 75% OR Quartile 2 1.45 1.24 1.07 1.46 1.28 1.02 1.37 1.19 1.18 p Value 0.33 0.58 0.87 0.33 0.52 0.95 0.42 0.65 0.71 Lower limit 0.688 0.587 0.466 0.686 0.606 0.445 0.636 0.559 0.500 of 95% CI Upper limit 3.07 2.60 2.46 3.11 2.71 2.36 2.95 2.55 2.78 of 95% CI OR Quartile 3 2.07 1.81 2.12 1.87 1.72 2.00 1.53 1.40 2.18 p Value 0.028 0.073 0.046 0.059 0.10 0.067 0.21 0.32 0.043 Lower limit 1.08 0.947 1.01 0.977 0.899 0.952 0.792 0.725 1.03 of 95% CI Upper limit 3.97 3.47 4.44 3.60 3.31 4.20 2.94 2.69 4.63 of 95% CI OR Quartile 4 1.22 1.24 0.933 1.22 1.20 0.976 0.986 0.971 1.02 p Value 0.60 0.57 0.87 0.60 0.63 0.95 0.97 0.94 0.96 Lower limit 0.582 0.593 0.406 0.583 0.573 0.424 0.465 0.458 0.443 of 95% CI Upper limit 2.54 2.59 2.14 2.55 2.51 2.25 2.09 2.06 2.36 of 95% CI

TABLE 20.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3930 4670 3930 4580 4090 4480 Average 4330 5030 4350 5030 4590 4770 Stdev 2660 3300 2640 3350 2860 3190 p (t-test) 0.15 0.17 0.71 Min 276 62.8 276 62.8 276 62.8 Max 11800 21200 11800 21200 13800 21200 n (Patient) 78 73 81 70 88 63 sCr only Median 3930 4670 3930 4580 4090 4490 Average 4340 5050 4360 5050 4510 4900 Stdev 2640 3340 2620 3390 2760 3330 p (t-test) 0.15 0.16 0.44 Min 276 62.8 276 62.8 276 62.8 Max 11800 21200 11800 21200 13800 21200 n (Patient) 79 71 82 68 88 62 UO only Median 3930 4790 3930 4790 3930 4800 Average 4500 5170 4500 5170 4470 5270 Stdev 2780 3480 2780 3480 2780 3460 p (t-test) 0.22 0.22 0.15 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 108 42 108 42 109 41 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.57 0.56 0.57 0.56 0.56 0.57 0.52 0.53 0.58 SE 0.047 0.047 0.053 0.047 0.047 0.053 0.048 0.048 0.053 p Value 0.16 0.17 0.21 0.20 0.22 0.21 0.66 0.51 0.12 nCohort 78 79 108 81 82 108 88 88 109 Non-persistent nCohort 73 71 42 70 68 42 63 62 41 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 78% 77% 76% 79% 78% 76% 78% 77% 78% Specificity 28% 28% 26% 28% 28% 26% 27% 27% 27% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 59% 58% 62% 59% 57% 62% 56% 55% 63% Specificity 58% 57% 55% 57% 56% 55% 53% 53% 55% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 27% 28% 24% 27% 28% 24% 24% 26% 24% Specificity 77% 77% 74% 77% 77% 74% 74% 75% 74% OR Quartile 2 1.40 1.33 1.12 1.45 1.38 1.12 1.31 1.29 1.29 p Value 0.37 0.46 0.79 0.33 0.40 0.79 0.48 0.52 0.56 Lower limit 0.666 0.631 0.488 0.688 0.651 0.488 0.616 0.603 0.549 of 95% CI Upper limit 2.94 2.79 2.57 3.07 2.91 2.57 2.80 2.74 3.02 of 95% CI OR Quartile 3 1.95 1.81 1.96 1.86 1.72 1.96 1.43 1.39 2.12 p Value 0.042 0.073 0.071 0.061 0.10 0.071 0.28 0.32 0.046 Lower limit 1.02 0.946 0.944 0.972 0.898 0.944 0.748 0.725 1.01 of 95% CI Upper limit 3.73 3.46 4.06 3.55 3.29 4.06 2.74 2.67 4.44 of 95% CI OR Quartile 4 1.26 1.33 0.893 1.22 1.29 0.893 0.883 1.04 0.933 p Value 0.54 0.45 0.79 0.60 0.50 0.79 0.75 0.91 0.87 Lower limit 0.602 0.636 0.389 0.582 0.615 0.389 0.417 0.495 0.406 of 95% CI Upper limit 2.63 2.78 2.05 2.54 2.69 2.05 1.87 2.20 2.14 of 95% CI

Example 21. Use of C—C Motif Chemokine 15 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 15 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 21.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.1 428 59.2 423 58.2 565 Average 764 2280 881 2540 859 2820 Stdev 4180 6070 3730 6700 3560 7110 p (t-test) 0.017 0.0070 0.0015 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 118 198 157 159 178 138 sCr only Median 22.1 428 59.2 434 57.2 596 Average 738 2400 843 2640 812 2980 Stdev 4000 6250 3680 6790 3510 7250 p (t-test) 0.0080 0.0036 5.0E−4 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 129 186 161 154 183 132 UO only Median 108 420 127 434 127 585 Average 1750 1620 1740 1630 1710 1780 Stdev 6070 2990 5930 3050 5860 3210 p (t-test) 0.86 0.88 0.94 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 18400 53500 18400 53500 18400 n (Patient) 240 74 255 59 262 52 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.73 0.61 0.67 0.68 0.59 0.69 0.71 0.61 SE 0.028 0.028 0.039 0.030 0.030 0.042 0.030 0.030 0.045 p Value 0 2.2E−16 0.0048 2.6E−8 1.5E−9 0.026 1.1E−10 1.5E−12 0.011 nCohort 118 129 240 157 161 255 178 183 262 Non-persistent nCohort 198 186 74 159 154 59 138 132 52 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 85% 85% 85% 84% 85% 81% 86% 86% 83% Specificity 37% 36% 26% 31% 32% 24% 30% 31% 24% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 65% 66% 65% 63% 64% 66% 67% 70% 69% Specificity 75% 72% 55% 63% 63% 54% 63% 64% 54% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 36% 37% 32% 36% 38% 32% 40% 42% 35% Specificity 93% 92% 77% 87% 87% 76% 87% 87% 77% OR Quartile 2 3.33 3.38 1.99 2.25 2.64 1.40 2.57 2.79 1.54 p Value 1.2E−5 1.1E−5 0.054 0.0032 5.9E−4 0.35 0.0012 6.2E−4 0.27 Lower limit 1.94 1.96 0.988 1.31 1.52 0.686 1.45 1.55 0.714 of 95% CI Upper limit 5.71 5.81 4.03 3.87 4.59 2.87 4.55 5.03 3.34 of 95% CI OR Quartile 3 5.36 4.92 2.22 2.89 3.11 2.26 3.59 4.08 2.62 p Value 8.1E−11 1.7E−10 0.0039 5.1E−6 1.3E−6 0.0069 8.9E−8 8.5E−9 0.0030 Lower limit 3.23 3.02 1.29 1.83 1.96 1.25 2.25 2.53 1.39 of 95% CI Upper limit 8.90 8.03 3.81 4.57 4.93 4.09 5.74 6.58 4.96 of 95% CI OR Quartile 4 7.69 7.02 1.61 3.72 4.03 1.54 4.25 5.13 1.74 p Value 2.4E−7 7.7E−8 0.10 4.6E−6 1.2E−6 0.17 2.3E−7 8.8E−9 0.088 Lower limit 3.54 3.45 0.911 2.12 2.29 0.832 2.46 2.94 0.921 of 95% CI Upper limit 16.7 14.3 2.86 6.52 7.07 2.86 7.36 8.95 3.31 of 95% CI

TABLE 21.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.0 423 53.2 436 54.2 565 Average 793 2220 857 2450 849 2650 Stdev 4300 5980 3850 6490 3700 6810 p (t-test) 0.027 0.0096 0.0034 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 111 205 146 170 164 152 sCr only Median 22.0 428 53.2 436 53.2 576 Average 750 2360 836 2550 830 2740 Stdev 4060 6190 3770 6630 3650 6910 p (t-test) 0.011 0.0053 0.0020 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 125 190 153 162 168 147 UO only Median 88.0 406 110 434 113 565 Average 1750 1650 1740 1680 1700 1790 Stdev 6210 3290 6030 3410 5950 3540 p (t-test) 0.87 0.93 0.91 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 221 93 237 77 244 70 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.73 0.61 0.69 0.70 0.60 0.70 0.71 0.61 SE 0.028 0.028 0.036 0.029 0.029 0.038 0.029 0.029 0.040 p Value 0 0 0.0025 1.2E−10 2.9E−11 0.0086 3.9E−12 3.2E−13 0.0038 nCohort 111 125 221 146 153 237 164 168 244 Non-persistent nCohort 205 190 93 170 162 77 152 147 70 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 84% 86% 84% 84% 86% 82% 85% 86% 83% Specificity 38% 38% 26% 32% 33% 25% 31% 32% 25% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 64% 65% 63% 64% 65% 65% 68% 69% 67% Specificity 76% 73% 56% 66% 65% 55% 66% 66% 55% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 35% 36% 31% 36% 38% 31% 39% 40% 33% Specificity 93% 92% 77% 88% 88% 77% 88% 88% 77% OR Quartile 2 3.29 3.64 1.85 2.51 3.02 1.49 2.53 3.01 1.61 p Value 1.4E−5 3.4E−6 0.055 7.9E−4 9.3E−5 0.23 0.0010 1.6E−4 0.17 Lower limit 1.92 2.11 0.987 1.47 1.74 0.779 1.46 1.70 0.811 of 95% CI Upper limit 5.64 6.27 3.47 4.31 5.26 2.86 4.40 5.33 3.20 of 95% CI OR Quartile 3 5.51 5.03 2.18 3.54 3.48 2.25 4.17 4.28 2.49 p Value 1.2E−10 1.5E−10 0.0022 1.0E−7 1.4E−7 0.0029 2.6E−9 1.7E−9 0.0014 Lower limit 3.28 3.07 1.32 2.22 2.19 1.32 2.60 2.66 1.42 of 95% CI Upper limit 9.25 8.24 3.59 5.63 5.53 3.84 6.66 6.86 4.35 of 95% CI OR Quartile 4 6.82 6.56 1.55 4.36 4.53 1.50 4.57 4.96 1.64 p Value 1.2E−6 2.1E−7 0.11 1.2E−6 4.3E−7 0.16 1.8E−7 4.0E−8 0.094 Lower limit 3.14 3.22 0.903 2.40 2.52 0.848 2.58 2.80 0.919 of 95% CI Upper limit 14.8 13.3 2.66 7.89 8.14 2.65 8.08 8.79 2.94 of 95% CI

TABLE 21.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.1 414 53.2 434 53.2 550 Average 800 2210 870 2420 866 2560 Stdev 4320 5970 3890 6440 3770 6670 p (t-test) 0.030 0.012 0.0060 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 110 206 143 173 157 159 sCr only Median 22.1 414 54.2 434 54.2 550 Average 762 2330 848 2510 855 2640 Stdev 4090 6160 3800 6580 3720 6790 p (t-test) 0.013 0.0070 0.0039 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 123 192 150 165 162 153 UO only Median 91.3 347 116 351 118 406 Average 1810 1540 1780 1570 1740 1660 Stdev 6340 3150 6180 3250 6100 3360 p (t-test) 0.67 0.76 0.91 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 211 103 224 90 231 83 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.72 0.59 0.68 0.69 0.58 0.70 0.70 0.59 SE 0.028 0.028 0.035 0.030 0.030 0.036 0.029 0.029 0.037 p Value 4.4E−16 1.3E−15 0.0084 6.1E−10 1.5E−10 0.029 2.2E−11 1.4E−11 0.016 nCohort 110 123 211 143 150 224 157 162 231 Non-persistent nCohort 206 192 103 173 165 90 159 153 83 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 84% 85% 83% 83% 85% 80% 84% 86% 81% Specificity 37% 37% 26% 31% 33% 25% 31% 32% 25% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 64% 65% 60% 64% 64% 60% 66% 67% 61% Specificity 75% 72% 55% 66% 65% 54% 66% 65% 54% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 34% 36% 31% 36% 37% 31% 38% 39% 33% Specificity 93% 92% 78% 88% 88% 77% 88% 88% 77% OR Quartile 2 3.12 3.24 1.66 2.28 2.72 1.30 2.43 2.81 1.37 p Value 3.3E−5 1.9E−5 0.093 0.0024 3.3E−4 0.39 0.0014 2.9E−4 0.32 Lower limit 1.82 1.89 0.919 1.34 1.57 0.715 1.41 1.61 0.736 of 95% CI Upper limit 5.33 5.56 3.02 3.88 4.69 2.37 4.19 4.92 2.56 of 95% CI OR Quartile 3 5.37 4.77 1.85 3.46 3.39 1.76 3.82 3.79 1.88 p Value 2.2E−10 5.4E−10 0.012 1.7E−7 2.4E−7 0.025 1.8E−8 2.3E−8 0.016 Lower limit 3.20 2.91 1.14 2.17 2.13 1.07 2.39 2.37 1.13 of 95% CI Upper limit 9.02 7.82 2.98 5.50 5.38 2.90 6.08 6.04 3.14 of 95% CI OR Quartile 4 6.71 6.34 1.57 4.14 4.30 1.53 4.40 4.46 1.66 p Value 1.5E−6 3.5E−7 0.093 2.8E−6 1.0E−6 0.12 4.8E−7 2.8E−7 0.073 Lower limit 3.09 3.11 0.927 2.29 2.40 0.889 2.47 2.52 0.954 of 95% CI Upper limit 14.6 12.9 2.67 7.50 7.72 2.64 7.84 7.88 2.89 of 95% CI

TABLE 21.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.0 406 53.2 434 53.2 537 Average 805 2200 875 2400 875 2530 Stdev 4340 5950 3910 6430 3800 6630 p (t-test) 0.032 0.014 0.0073 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 109 207 142 174 155 161 sCr only Median 22.1 414 53.2 434 53.2 544 Average 762 2330 853 2500 860 2620 Stdev 4090 6160 3820 6560 3730 6770 p (t-test) 0.013 0.0077 0.0043 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 123 192 149 166 161 154 UO only Median 88.0 338 115 338 115 341 Average 1850 1480 1820 1500 1790 1560 Stdev 6400 3110 6250 3180 6170 3270 p (t-test) 0.58 0.64 0.75 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 207 107 219 95 225 89 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.72 0.59 0.68 0.69 0.58 0.70 0.70 0.58 SE 0.028 0.028 0.034 0.030 0.030 0.036 0.029 0.029 0.036 p Value 2.2E−16 1.3E−15 0.011 6.8E−10 1.7E−10 0.035 2.4E−11 1.6E−11 0.021 nCohort 109 123 207 142 149 219 155 161 225 Non-persistent nCohort 207 192 107 174 166 95 161 154 89 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 84% 85% 83% 83% 85% 81% 84% 86% 82% Specificity 38% 37% 27% 32% 33% 25% 32% 32% 25% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 64% 65% 60% 63% 64% 60% 66% 66% 62% Specificity 76% 72% 55% 66% 65% 54% 66% 65% 55% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 34% 36% 30% 36% 37% 29% 37% 38% 30% Specificity 93% 92% 77% 88% 88% 77% 88% 88% 77% OR Quartile 2 3.18 3.24 1.79 2.32 2.76 1.43 2.51 2.86 1.55 p Value 2.5E−5 1.9E−5 0.055 0.0020 2.6E−4 0.24 9.0E−4 2.3E−4 0.17 Lower limit 1.86 1.89 0.989 1.36 1.60 0.790 1.46 1.64 0.834 of 95% CI Upper limit 5.44 5.56 3.24 3.95 4.77 2.61 4.34 5.01 2.87 of 95% CI OR Quartile 3 5.62 4.77 1.82 3.37 3.30 1.78 3.82 3.68 1.95 p Value 1.1E−10 5.4E−10 0.013 3.1E−7 4.3E−7 0.020 1.8E−8 4.2E−8 0.0090 Lower limit 3.33 2.91 1.14 2.12 2.08 1.09 2.40 2.31 1.18 of 95% CI Upper limit 9.49 7.82 2.93 5.36 5.23 2.91 6.08 5.86 3.22 of 95% CI OR Quartile 4 6.59 6.34 1.45 4.07 4.23 1.38 4.25 4.38 1.45 p Value 1.9E−6 3.5E−7 0.16 3.6E−6 1.4E−6 0.25 8.7E−7 3.8E−7 0.18 Lower limit 3.04 3.11 0.858 2.25 2.36 0.801 2.39 2.48 0.838 of 95% CI Upper limit 14.3 12.9 2.46 7.37 7.59 2.36 7.57 7.74 2.51 of 95% CI

TABLE 21.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent'’ and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 22.0 406 53.2 434 53.2 497 Average 799 2170 889 2350 893 2450 Stdev 4420 5900 3970 6340 3870 6530 p (t-test) 0.036 0.019 0.012 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 105 211 137 179 149 167 sCr only Median 22.1 423 53.2 436 53.2 544 Average 749 2320 836 2470 845 2590 Stdev 4130 6120 3860 6490 3770 6690 p (t-test) 0.014 0.0082 0.0047 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 34200 53500 34200 53500 34200 53500 n (Patient) 120 195 145 170 157 158 UO only Median 81.4 347 88.0 341 97.4 344 Average 1830 1530 1820 1530 1790 1570 Stdev 6480 3070 6350 3120 6280 3180 p (t-test) 0.65 0.66 0.74 Min 0.0116 0.0116 0.0116 0.0116 0.0116 0.0116 Max 53500 19200 53500 19200 53500 19200 n (Patient) 201 113 211 103 216 98 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.73 0.61 0.68 0.70 0.59 0.69 0.71 0.59 SE 0.028 0.028 0.034 0.030 0.029 0.035 0.029 0.029 0.035 p Value 0 2.2E−16 0.0014 1.1E−9 2.1E−11 0.0075 4.6E−11 1.9E−12 0.0076 nCohort 105 120 201 137 145 211 149 157 216 Non-persistent nCohort 211 195 113 179 170 103 167 158 98 Persistent Cutoff 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 0.0489 Quartile 2 Sensitivity 84% 85% 84% 83% 85% 83% 84% 85% 83% Specificity 38% 37% 27% 32% 33% 26% 32% 32% 26% Cutoff 155 154 155 155 154 155 155 154 155 Quartile 3 Sensitivity 64% 65% 62% 63% 64% 61% 65% 66% 62% Specificity 77% 73% 57% 66% 66% 55% 66% 66% 56% Cutoff 909 912 902 909 912 902 909 912 902 Quartile 4 Sensitivity 34% 36% 32% 35% 37% 31% 37% 39% 32% Specificity 93% 92% 79% 88% 89% 78% 88% 89% 78% OR Quartile 2 3.20 3.18 1.99 2.35 2.74 1.66 2.58 2.82 1.67 p Value 2.2E−5 2.4E−5 0.023 0.0016 2.7E−4 0.093 6.1E−4 2.4E−4 0.098 Lower limit 1.87 1.86 1.10 1.38 1.59 0.919 1.50 1.62 0.911 of 95% CI Upper limit 5.49 5.45 3.59 4.00 4.71 3.02 4.43 4.91 3.05 of 95% CI OR Quartile 3 5.87 5.02 2.13 3.31 3.50 1.96 3.62 3.89 2.06 p Value 8.7E−11 2.5E−10 0.0016 5.0E−7 1.3E−7 0.0060 5.7E−8 1.2E−8 0.0037 Lower limit 3.44 3.05 1.33 2.07 2.20 1.21 2.28 2.44 1.26 of 95% CI Upper limit 10.0 8.28 3.42 5.27 5.58 3.17 5.77 6.20 3.36 of 95% CI OR Quartile 4 7.25 6.91 1.72 4.11 4.75 1.57 4.19 4.86 1.62 p Value 2.1E−6 3.0E−7 0.041 4.7E−6 4.6E−7 0.093 1.6E−6 1.3E−7 0.076 Lower limit 3.20 3.30 1.02 2.24 2.59 0.927 2.33 2.70 0.950 of 95% CI Upper limit 16.4 14.5 2.89 7.52 8.70 2.67 7.52 8.73 2.76 of 95% CI

TABLE 21.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3500 4670 3940 4690 4060 4580 Average 3630 5180 4270 5060 4340 5090 Stdev 2250 3190 2560 3340 2610 3410 p (t-test) 0.0024 0.10 0.13 Min 62.8 413 62.8 413 62.8 413 Max 10700 21200 11800 21200 11800 21200 n (Patient) 50 101 75 76 85 66 sCr only Median 3700 4680 3950 4670 3950 4720 Average 3750 5170 4340 4990 4300 5170 Stdev 2150 3280 2490 3410 2460 3570 p (t-test) 0.0056 0.19 0.079 Min 276 62.8 276 62.8 276 62.8 Max 10700 21200 11800 21200 11800 21200 n (Patient) 52 98 73 77 86 64 UO only Median 4050 4720 4050 4840 4120 4800 Average 4550 5100 4490 5500 4500 5660 Stdev 2770 3580 2700 3890 2680 4160 p (t-test) 0.33 0.096 0.076 Min 62.8 1120 62.8 1120 62.8 1630 Max 13800 21200 13800 21200 13800 21200 n (Patient) 112 38 120 30 125 25 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.63 0.54 0.56 0.55 0.58 0.56 0.57 0.58 SE 0.046 0.046 0.055 0.047 0.047 0.060 0.047 0.048 0.065 p Value 8.4E−4 0.0045 0.44 0.17 0.32 0.16 0.24 0.17 0.22 nCohort 50 52 112 75 73 120 85 86 125 Non-persistent nCohort 101 98 38 76 77 30 66 64 25 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 79% 78% 76% 75% 73% 80% 76% 77% 80% Specificity 34% 31% 26% 25% 23% 27% 26% 27% 26% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 58% 57% 58% 57% 56% 67% 56% 58% 64% Specificity 66% 63% 53% 56% 56% 54% 54% 56% 53% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 32% 33% 24% 29% 30% 30% 29% 31% 28% Specificity 88% 88% 74% 79% 79% 76% 78% 79% 75% OR Quartile 2 1.96 1.54 1.13 1.02 0.810 1.45 1.09 1.19 1.43 p Value 0.081 0.27 0.79 0.96 0.58 0.45 0.82 0.65 0.50 Lower limit 0.920 0.721 0.477 0.488 0.387 0.545 0.519 0.563 0.498 of 95% CI 4.18 3.27 2.66 2.12 1.70 3.88 2.29 2.52 4.13 Upper limit of 95% CI OR Quartile 3 2.73 2.32 1.53 1.66 1.62 2.36 1.50 1.73 1.99 p Value 0.0054 0.017 0.26 0.12 0.14 0.045 0.22 0.100 0.13 Lower limit 1.35 1.16 0.728 0.872 0.850 1.02 0.788 0.900 0.818 of 95% CI Upper limit 5.53 4.63 3.22 3.16 3.09 5.47 2.87 3.33 4.84 of 95% CI OR Quartile 4 3.40 3.72 0.888 1.50 1.65 1.34 1.40 1.72 1.18 p Value 0.012 0.0067 0.79 0.28 0.19 0.51 0.37 0.15 0.74 Lower limit 1.31 1.44 0.376 0.715 0.779 0.555 0.671 0.818 0.450 of 95% CI Upper limit 8.80 9.61 2.10 3.16 3.48 3.26 2.94 3.60 3.09 of 95% CI

TABLE 21.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3620 4600 3930 4500 3940 4490 Average 3690 5130 4340 4950 4400 4960 Stdev 2220 3210 2550 3320 2610 3360 p (t-test) 0.0053 0.21 0.26 Min 62.8 276 62.8 276 62.8 276 Max 10700 21200 11800 21200 11800 21200 n (Patient) 49 102 70 81 79 72 sCr only Median 3730 4670 4000 4490 3940 4500 Average 3820 5120 4410 4910 4350 5030 Stdev 2120 3300 2500 3390 2480 3480 p (t-test) 0.012 0.31 0.17 Min 703 62.8 703 62.8 703 62.8 Max 10700 21200 11800 21200 11800 21200 n (Patient) 51 99 70 80 79 71 UO only Median 3940 4710 3940 4840 4050 4790 Average 4500 5110 4440 5420 4450 5520 Stdev 2770 3420 2700 3640 2680 3820 p (t-test) 0.25 0.078 0.067 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 103 47 111 39 116 34 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.62 0.56 0.55 0.53 0.59 0.54 0.55 0.59 SE 0.046 0.047 0.051 0.047 0.047 0.054 0.047 0.047 0.057 p Value 0.0027 0.012 0.27 0.30 0.48 0.089 0.38 0.31 0.13 nCohort 49 51 103 70 70 111 79 79 116 Non-persistent nCohort 102 99 47 81 80 39 72 71 34 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 78% 77% 77% 74% 72% 79% 75% 75% 79% Specificity 33% 29% 26% 24% 23% 27% 25% 25% 27% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 58% 57% 60% 57% 55% 67% 57% 56% 65% Specificity 65% 63% 54% 57% 56% 56% 56% 56% 54% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 31% 32% 26% 28% 29% 31% 28% 30% 29% Specificity 88% 88% 75% 79% 79% 77% 77% 78% 76% OR Quartile 2 1.76 1.38 1.16 0.916 0.781 1.44 1.02 0.998 1.41 p Value 0.14 0.41 0.71 0.82 0.51 0.42 0.96 1.00 0.47 Lower limit 0.824 0.643 0.520 0.438 0.372 0.594 0.487 0.478 0.556 of 95% CI Upper limit 3.77 2.95 2.60 1.92 1.64 3.47 2.12 2.09 3.56 of 95% CI OR Quartile 3 2.58 2.19 1.76 1.75 1.54 2.53 1.66 1.62 2.18 p Value 0.0086 0.026 0.11 0.089 0.19 0.017 0.12 0.14 0.054 Lower limit 1.27 1.10 0.872 0.918 0.806 1.18 0.873 0.850 0.986 of 95% CI Upper limit 5.24 4.39 3.54 3.34 2.93 5.43 3.17 3.09 4.81 of 95% CI OR Quartile 4 3.28 3.58 1.02 1.45 1.48 1.45 1.30 1.53 1.31 p Value 0.014 0.0085 0.97 0.33 0.31 0.37 0.48 0.26 0.53 Lower limit 1.27 1.38 0.460 0.688 0.700 0.647 0.624 0.731 0.559 of 95% CI Upper limit 8.48 9.26 2.24 3.07 3.13 3.26 2.72 3.21 3.07 of 95% CI

TABLE 21.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3620 4600 3930 4580 3920 4690 Average 3690 5130 4300 4980 4240 5090 Stdev 2220 3210 2550 3310 2530 3360 p (t-test) 0.0053 0.17 0.081 Min 62.8 276 62.8 276 62.8 276 Max 10700 21200 11800 21200 11800 21200 n (Patient) 49 102 69 82 75 76 sCr only Median 3730 4670 3940 4500 3930 4670 Average 3820 5120 4370 4930 4300 5040 Stdev 2120 3300 2500 3370 2490 3430 p (t-test) 0.012 0.25 0.13 Min 703 62.8 703 62.8 703 62.8 Max 10700 21200 11800 21200 11800 21200 n (Patient) 51 99 69 81 75 75 UO only Median 3930 4790 3920 4860 3930 4910 Average 4400 5250 4320 5550 4340 5650 Stdev 2720 3410 2660 3550 2630 3690 p (t-test) 0.098 0.021 0.018 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 98 52 105 45 110 40 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.62 0.58 0.56 0.54 0.62 0.58 0.56 0.62 SE 0.046 0.047 0.050 0.047 0.047 0.051 0.046 0.047 0.054 p Value 0.0027 0.012 0.096 0.22 0.37 0.017 0.10 0.20 0.027 nCohort 49 51 98 69 69 105 75 75 110 Non-persistent nCohort 102 99 52 82 81 45 76 75 40 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 78% 77% 77% 74% 73% 80% 76% 75% 80% Specificity 33% 29% 27% 25% 23% 28% 27% 25% 27% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 58% 57% 62% 57% 56% 69% 59% 57% 68% Specificity 65% 63% 56% 58% 57% 58% 59% 57% 56% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 31% 32% 29% 29% 30% 33% 30% 31% 32% Specificity 88% 88% 77% 80% 80% 78% 80% 80% 77% OR Quartile 2 1.76 1.38 1.20 0.950 0.810 1.53 1.17 1.00 1.50 p Value 0.14 0.41 0.64 0.89 0.58 0.33 0.67 1.0 0.37 Lower limit 0.824 0.643 0.549 0.454 0.385 0.655 0.561 0.479 0.621 of 95% CI Upper limit 3.77 2.95 2.64 1.99 1.70 3.56 2.45 2.09 3.62 of 95% CI OR Quartile 3 2.58 2.19 2.05 1.85 1.62 3.07 2.06 1.81 2.68 p Value 0.0086 0.026 0.041 0.062 0.14 0.0030 0.029 0.073 0.011 Lower limit 1.27 1.10 1.03 0.969 0.851 1.46 1.08 0.945 1.25 of 95% CI Upper limit 5.24 4.39 4.07 3.54 3.10 6.44 3.94 3.45 5.74 of 95% CI OR Quartile 4 3.28 3.58 1.32 1.63 1.65 1.78 1.74 1.77 1.64 p Value 0.014 0.0085 0.47 0.21 0.19 0.14 0.15 0.14 0.23 Lower limit 1.27 1.38 0.618 0.764 0.777 0.823 0.822 0.837 0.737 of 95% CI Upper limit 8.48 9.26 2.83 3.46 3.52 3.86 3.67 3.74 3.64 of 95% CI

TABLE 21.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3620 4600 3930 4670 3860 4710 Average 3690 5130 4280 4980 4220 5100 Stdev 2220 3210 2560 3290 2540 3340 p (t-test) 0.0053 0.15 0.072 Min 62.8 276 62.8 276 62.8 276 Max 10700 21200 11800 21200 11800 21200 n (Patient) 49 102 68 83 74 77 sCr only Median 3730 4670 3930 4580 3930 4690 Average 3820 5120 4350 4940 4290 5050 Stdev 2120 3300 2510 3350 2500 3400 p (t-test) 0.012 0.23 0.12 Min 703 62.8 703 62.8 703 62.8 Max 10700 21200 11800 21200 11800 21200 n (Patient) 51 99 68 82 74 76 UO only Median 3920 4800 3920 4860 3930 4860 Average 4390 5200 4340 5400 4360 5470 Stdev 2750 3330 2710 3410 2680 3530 p (t-test) 0.11 0.041 0.037 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 95 55 100 50 105 45 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.62 0.58 0.56 0.55 0.61 0.58 0.57 0.60 SE 0.046 0.047 0.049 0.047 0.047 0.050 0.046 0.047 0.052 p Value 0.0027 0.012 0.089 0.17 0.31 0.027 0.081 0.16 0.043 nCohort 49 51 95 68 68 100 74 74 105 Non-persistent nCohort 102 99 55 83 82 50 77 76 45 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 78% 77% 76% 75% 73% 78% 77% 75% 78% Specificity 33% 29% 26% 25% 24% 27% 27% 26% 27% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 58% 57% 62% 58% 56% 66% 60% 58% 64% Specificity 65% 63% 57% 59% 57% 58% 59% 58% 56% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 31% 32% 27% 29% 29% 30% 30% 30% 29% Specificity 88% 88% 76% 79% 79% 77% 80% 80% 76% OR Quartile 2 1.76 1.38 1.15 0.984 0.839 1.31 1.21 1.04 1.27 p Value 0.14 0.41 0.72 0.97 0.64 0.51 0.61 0.92 0.57 Lower limit 0.824 0.643 0.533 0.470 0.399 0.588 0.582 0.496 0.558 of 95% CI Upper limit 3.77 2.95 2.50 2.06 1.76 2.92 2.53 2.16 2.90 of 95% CI OR Quartile 3 2.58 2.19 2.13 1.96 1.72 2.68 2.18 1.91 2.32 p Value 0.0086 0.026 0.029 0.043 0.10 0.0063 0.019 0.051 0.022 Lower limit 1.27 1.10 1.08 1.02 0.898 1.32 1.14 0.997 1.13 of 95% CI Upper limit 5.24 4.39 4.20 3.75 3.29 5.44 4.17 3.65 4.79 of 95% CI OR Quartile 4 3.28 3.58 1.17 1.57 1.60 1.43 1.68 1.71 1.30 p Value 0.014 0.0085 0.68 0.24 0.23 0.35 0.18 0.16 0.51 Lower limit 1.27 1.38 0.551 0.737 0.749 0.669 0.793 0.807 0.593 of 95% CI Upper limit 8.48 9.26 2.50 3.34 3.40 3.08 3.54 3.61 2.85 of 95% CI

TABLE 21.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 3620 4600 3930 4490 3930 4500 Average 3740 5090 4360 4890 4340 4950 Stdev 2190 3220 2560 3280 2530 3340 p (t-test) 0.0100 0.28 0.21 Min 703 62.8 703 62.8 703 62.8 Max 10700 21200 11800 21200 11800 21200 n (Patient) 47 104 65 86 70 81 sCr only Median 3700 4680 3930 4580 3930 4690 Average 3790 5120 4360 4920 4290 5030 Stdev 2130 3280 2540 3320 2520 3370 p (t-test) 0.010 0.26 0.14 Min 703 62.8 703 62.8 703 62.8 Max 10700 21200 11800 21200 11800 21200 n (Patient) 50 100 66 84 72 78 UO only Median 3930 4750 3920 4800 3930 4780 Average 4430 5100 4410 5180 4420 5220 Stdev 2790 3280 2750 3340 2730 3410 p (t-test) 0.19 0.13 0.12 Min 62.8 513 62.8 513 62.8 513 Max 13800 21200 13800 21200 13800 21200 n (Patient) 92 58 95 55 99 51 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.62 0.57 0.55 0.55 0.58 0.55 0.56 0.57 SE 0.047 0.047 0.049 0.047 0.047 0.049 0.047 0.047 0.050 p Value 0.0049 0.0079 0.17 0.34 0.32 0.11 0.27 0.17 0.14 nCohort 47 50 92 65 66 95 70 72 99 Non-persistent nCohort 104 100 58 86 84 55 81 78 51 Persistent Cutoff 2740 2730 2740 2740 2730 2740 2740 2730 2740 Quartile 2 Sensitivity 78% 77% 76% 74% 74% 76% 75% 76% 76% Specificity 32% 30% 26% 25% 24% 26% 26% 26% 26% Cutoff 4230 4260 4260 4230 4260 4260 4230 4260 4260 Quartile 3 Sensitivity 58% 57% 59% 57% 56% 62% 58% 58% 61% Specificity 66% 64% 55% 58% 58% 57% 59% 58% 56% Cutoff 6010 6060 6060 6010 6060 6060 6010 6060 6060 Quartile 4 Sensitivity 31% 32% 26% 28% 29% 27% 28% 29% 25% Specificity 87% 88% 75% 78% 79% 76% 79% 79% 75% OR Quartile 2 1.65 1.43 1.11 0.950 0.902 1.15 1.06 1.11 1.16 p Value 0.20 0.35 0.79 0.89 0.79 0.72 0.89 0.78 0.72 Lower limit 0.765 0.669 0.519 0.452 0.429 0.533 0.506 0.533 0.527 of 95% CI Upper limit 3.56 3.08 2.37 2.00 1.90 2.50 2.20 2.32 2.54 of 95% CI OR Quartile 3 2.64 2.36 1.76 1.86 1.72 2.13 1.95 1.91 1.94 p Value 0.0080 0.016 0.095 0.061 0.10 0.029 0.043 0.051 0.059 Lower limit 1.29 1.17 0.906 0.971 0.899 1.08 1.02 0.998 0.974 of 95% CI Upper limit 5.42 4.75 3.43 3.58 3.31 4.20 3.74 3.65 3.85 of 95% CI OR Quartile 4 3.04 3.45 1.05 1.41 1.49 1.17 1.45 1.59 1.01 p Value 0.022 0.011 0.91 0.37 0.31 0.68 0.33 0.23 0.97 Lower limit 1.17 1.33 0.492 0.662 0.697 0.551 0.688 0.752 0.466 of 95% CI Upper limit 7.87 8.93 2.22 3.00 3.17 2.50 3.07 3.36 2.20 of 95% CI

Example 22. Use of C—C Motif Chemokine 18 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 18 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 22.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.870 1.22 0.716 1.23 0.716 1.18 Average 0.858 2.90 0.745 2.86 0.739 2.83 Stdev 0.658 9.70 0.561 9.57 0.588 9.49 p (t-test) 0.32 0.33 0.35 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 2.61 60.0 2.01 60.0 2.01 60.0 n (Patient) 23 109 20 112 18 114 sCr only Median 0.797 1.27 0.819 1.26 0.797 1.24 Average 0.783 3.07 0.794 3.04 0.786 3.02 Stdev 0.513 10.1 0.519 10.0 0.527 9.96 p (t-test) 0.22 0.23 0.24 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 2.01 60.0 2.01 60.0 2.01 60.0 n (Patient) 30 101 29 102 28 103 UO only Median 0.970 1.36 0.947 1.42 0.915 1.36 Average 1.06 5.16 0.957 4.78 0.923 4.33 Stdev 0.834 14.3 0.720 13.4 0.707 12.6 p (t-test) 0.010 0.014 0.028 Min 0.00242 0.273 0.00242 0.273 0.00242 0.273 Max 4.23 60.0 3.20 60.0 3.20 60.0 n (Patient) 83 48 76 55 68 63 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.69 0.67 0.70 0.68 0.71 0.70 0.68 0.71 SE 0.058 0.051 0.050 0.057 0.052 0.047 0.059 0.052 0.045 p Value 0.0071 2.4E−4 8.4E−4 5.7E−4 5.5E−4 4.6E−6 9.9E−4 5.0E−4 4.7E−6 nCohort 23 30 83 20 29 76 18 28 68 Recovered nCohort 109 101 48 112 102 55 114 103 63 Non-recovered Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 77% 78% 90% 78% 77% 91% 78% 78% 89% Specificity 35% 37% 34% 40% 34% 37% 44% 36% 38% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 54% 56% 67% 54% 56% 69% 54% 55% 65% Specificity 70% 70% 59% 75% 69% 63% 72% 68% 63% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 28% 31% 31% 28% 30% 36% 27% 30% 35% Specificity 87% 93% 78% 90% 93% 83% 89% 93% 84% OR Quartile 2 1.79 2.08 4.38 2.32 1.81 5.83 2.85 1.93 4.95 p Value 0.24 0.10 0.0050 0.099 0.20 8.0E−4 0.046 0.15 7.0E−4 Lower limit 0.681 0.862 1.56 0.854 0.738 2.08 1.02 0.784 1.96 of 95% CI Upper limit 4.71 5.01 12.3 6.30 4.43 16.4 7.98 4.76 12.5 of 95% CI OR Quartile 3 2.70 3.02 2.88 3.59 2.81 3.83 2.99 2.62 3.21 p Value 0.044 0.013 0.0052 0.020 0.021 3.6E−4 0.050 0.033 0.0014 Lower limit 1.03 1.26 1.37 1.22 1.17 1.83 1.00 1.08 1.57 of 95% CI Upper limit 7.08 7.25 6.06 10.5 6.78 8.01 8.95 6.33 6.55 of 95% CI OR Quartile 4 2.53 6.20 1.64 3.44 5.89 2.77 2.99 5.60 2.78 p Value 0.16 0.017 0.23 0.11 0.020 0.014 0.16 0.024 0.015 Lower limit 0.701 1.39 0.735 0.755 1.32 1.23 0.649 1.25 1.22 of 95% CI Upper limit 9.14 27.7 3.66 15.7 26.3 6.23 13.8 25.1 6.36 of 95% CI

TABLE 22.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.695 1.32 0.659 1.32 0.604 1.30 Average 0.763 3.18 0.738 3.14 0.692 3.09 Stdev 0.593 10.2 0.599 10.1 0.577 10.0 p (t-test) 0.17 0.18 0.19 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 2.01 60.0 2.01 60.0 2.01 60.0 n (Patient) 35 97 33 99 30 102 sCr only Median 0.735 1.35 0.774 1.35 0.735 1.34 Average 0.781 3.36 0.788 3.33 0.782 3.31 Stdev 0.564 10.6 0.569 10.6 0.575 10.5 p (t-test) 0.12 0.13 0.13 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 2.01 60.0 2.01 60.0 2.01 60.0 n (Patient) 42 90 41 91 40 92 UO only Median 0.978 1.39 0.974 1.42 0.983 1.36 Average 1.08 5.69 1.06 5.25 1.04 4.66 Stdev 0.846 15.3 0.838 14.5 0.843 13.4 p (t-test) 0.0051 0.0091 0.020 Min 0.00242 0.273 0.00242 0.273 0.00242 0.273 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 89 42 84 47 76 55 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.72 0.67 0.73 0.71 0.68 0.74 0.71 0.67 SE 0.047 0.045 0.052 0.046 0.046 0.050 0.046 0.046 0.049 p Value 3.6E−6 1.2E−6 0.0010 1.0E−6 3.6E−6 4.4E−4 1.2E−7 3.3E−6 5.8E−4 nCohort 35 42 89 33 41 84 30 40 76 Recovered nCohort 97 90 42 99 91 47 102 92 55 Non-recovered Cutoff 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Quartile 2 Sensitivity 81% 82% 88% 82% 81% 89% 82% 82% 87% Specificity 43% 40% 31% 45% 39% 33% 50% 40% 34% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 58% 59% 69% 58% 58% 68% 57% 58% 64% Specificity 71% 69% 58% 73% 68% 60% 73% 68% 59% Cutoff 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Quartile 4 Sensitivity 30% 32% 33% 29% 32% 34% 29% 32% 33% Specificity 89% 90% 79% 88% 90% 80% 90% 90% 80% OR Quartile 2 3.29 3.14 3.40 3.75 2.79 4.20 4.67 2.94 3.57 p Value 0.0056 0.0061 0.021 0.0024 0.014 0.0064 5.9E−4 0.010 0.0070 Lower limit 1.42 1.39 1.21 1.60 1.23 1.50 1.94 1.29 1.42 of 95% CI Upper limit 7.65 7.14 9.57 8.81 6.32 11.8 11.2 6.70 8.98 of 95% CI OR Quartile 3 3.41 3.20 3.14 3.62 3.00 3.14 3.62 2.82 2.54 p Value 0.0040 0.0034 0.0040 0.0035 0.0056 0.0029 0.0050 0.0091 0.011 Lower limit 1.48 1.47 1.44 1.53 1.38 1.48 1.48 1.29 1.24 of 95% CI Upper limit 7.88 6.95 6.83 8.58 6.54 6.66 8.91 6.16 5.19 of 95% CI OR Quartile 4 3.31 4.52 1.84 3.00 4.33 2.03 3.75 4.14 1.98 p Value 0.038 0.0084 0.14 0.057 0.010 0.084 0.041 0.013 0.094 Lower limit 1.07 1.47 0.813 0.969 1.41 0.910 1.06 1.35 0.891 of 95% CI Upper limit 10.2 13.9 4.17 9.31 13.3 4.55 13.3 12.7 4.39 of 95% CI

TABLE 22.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.739 1.30 0.700 1.30 0.700 1.28 Average 0.865 3.50 0.851 3.45 0.842 3.34 Stdev 0.690 11.0 0.700 10.9 0.702 10.6 p (t-test) 0.100 0.11 0.13 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 3.02 60.0 3.02 60.0 3.02 60.0 n (Patient) 48 84 46 86 42 90 sCr only Median 0.797 1.34 0.819 1.33 0.819 1.28 Average 0.857 3.71 0.864 3.67 0.874 3.59 Stdev 0.614 11.4 0.617 11.3 0.622 11.2 p (t-test) 0.068 0.074 0.086 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 2.07 60.0 2.07 60.0 2.07 60.0 n (Patient) 54 78 53 79 51 81 UO only Median 1.02 1.36 1.00 1.36 1.00 1.36 Average 1.11 5.75 1.10 5.36 1.07 4.76 Stdev 0.842 15.5 0.855 14.8 0.863 13.7 p (t-test) 0.0050 0.0085 0.019 Min 0.00242 0.273 0.00242 0.273 0.00242 0.273 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 90 41 86 45 78 53 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.69 0.64 0.69 0.69 0.65 0.69 0.68 0.65 SE 0.046 0.045 0.054 0.046 0.046 0.052 0.047 0.046 0.050 p Value 4.4E−5 1.7E−5 0.010 2.5E−5 4.2E−5 0.0047 5.0E−5 1.6E−4 0.0023 nCohort Recovered 48 54 90 46 53 86 42 51 78 nCohort Non- 84 78 41 86 79 45 90 81 53 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Sensitivity 83% 83% 85% 84% 82% 87% 83% 81% 87% Specificity 40% 37% 30% 41% 36% 31% 43% 35% 33% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 58% 59% 63% 58% 58% 64% 57% 57% 62% Specificity 65% 63% 56% 65% 62% 57% 64% 61% 58% Cutoff Quartile 4 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Sensitivity 32% 35% 32% 31% 34% 31% 31% 33% 32% Specificity 88% 89% 78% 87% 89% 78% 88% 88% 79% OR Quartile 2 3.28 2.94 2.50 3.62 2.59 2.97 3.75 2.40 3.29 p Value 0.0043 0.0092 0.066 0.0021 0.020 0.028 0.0017 0.033 0.012 Lower limit of 95% CI 1.45 1.31 0.942 1.59 1.16 1.12 1.64 1.08 1.30 Upper limit of 95% CI 7.40 6.63 6.64 8.22 5.81 7.87 8.56 5.36 8.28 OR Quartile 3 2.55 2.44 2.17 2.60 2.30 2.40 2.35 2.04 2.25 p Value 0.012 0.014 0.046 0.012 0.022 0.021 0.027 0.051 0.026 Lower limit of 95% CI 1.23 1.20 1.01 1.24 1.13 1.14 1.10 0.998 1.10 Upper limit of 95% CI 5.32 4.99 4.63 5.47 4.69 5.06 5.02 4.16 4.60 OR Quartile 4 3.32 4.24 1.62 3.05 4.07 1.59 3.34 3.75 1.83 p Value 0.015 0.0035 0.25 0.024 0.0045 0.26 0.022 0.0075 0.14 Lower limit of 95% CI 1.26 1.61 0.713 1.15 1.54 0.708 1.19 1.42 0.825 Upper limit of 95% CI 8.75 11.2 3.71 8.06 10.7 3.58 9.41 9.88 4.06

TABLE 22.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.774 1.35 0.704 1.33 0.700 1.30 Average 0.862 3.74 0.846 3.61 0.837 3.45 Stdev 0.661 11.4 0.678 11.2 0.684 10.9 p (t-test) 0.065 0.080 0.11 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 3.02 60.0 3.02 60.0 3.02 60.0 n (Patient) 55 77 51 81 46 86 sCr only Median 0.774 1.36 0.797 1.35 0.819 1.35 Average 0.842 3.83 0.849 3.79 0.861 3.74 Stdev 0.606 11.6 0.609 11.5 0.608 11.4 p (t-test) 0.054 0.059 0.065 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 2.07 60.0 2.07 60.0 2.07 60.0 n (Patient) 57 75 56 76 55 77 UO only Median 1.02 1.36 1.00 1.36 1.02 1.35 Average 1.09 5.57 1.08 5.20 1.08 4.67 Stdev 0.825 15.1 0.840 14.5 0.864 13.6 p (t-test) 0.0063 0.010 0.022 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 88 43 84 47 77 54 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.71 0.64 0.71 0.70 0.65 0.70 0.69 0.64 SE 0.045 0.044 0.053 0.045 0.045 0.051 0.046 0.045 0.050 p Value 5.3E−6 1.6E−6 0.0064 4.1E−6 4.5E−6 0.0045 1.4E−5 2.0E−5 0.0056 nCohort Recovered 55 57 88 51 56 84 46 55 77 nCohort Non- 77 75 43 81 76 47 86 77 54 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Sensitivity 84% 85% 84% 85% 84% 85% 85% 83% 85% Specificity 38% 39% 30% 41% 38% 31% 43% 36% 32% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 61% 61% 63% 60% 61% 64% 58% 60% 61% Specificity 65% 65% 56% 67% 64% 57% 65% 64% 57% Cutoff Quartile 4 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Sensitivity 35% 36% 35% 33% 36% 32% 33% 35% 31% Specificity 89% 89% 80% 88% 89% 79% 89% 89% 79% OR Quartile 2 3.35 3.66 2.16 4.02 3.20 2.56 4.32 2.81 2.76 p Value 0.0040 0.0023 0.11 9.9E−4 0.0054 0.047 5.5E−4 0.012 0.025 Lower limit of 95% CI 1.47 1.59 0.851 1.76 1.41 1.01 1.88 1.25 1.14 Upper limit of 95% CI 7.61 8.41 5.47 9.22 7.27 6.47 9.90 6.33 6.73 OR Quartile 3 2.97 2.93 2.12 3.06 2.76 2.35 2.60 2.60 2.10 p Value 0.0031 0.0032 0.049 0.0028 0.0053 0.023 0.012 0.0088 0.041 Lower limit of 95% CI 1.44 1.43 1.00 1.47 1.35 1.13 1.24 1.27 1.03 Upper limit of 95% CI 6.10 6.00 4.48 6.37 5.64 4.91 5.47 5.30 4.26 OR Quartile 4 4.41 4.78 2.08 3.75 4.59 1.72 3.96 4.41 1.75 p Value 0.0027 0.0015 0.077 0.0075 0.0020 0.19 0.0090 0.0027 0.17 Lower limit of 95% CI 1.67 1.82 0.924 1.42 1.74 0.769 1.41 1.67 0.791 Upper limit of 95% CI 11.6 12.6 4.70 9.88 12.1 3.84 11.1 11.6 3.88

TABLE 22.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 1.02 1.35 1.02 1.33 1.02 1.33 Average 1.04 4.93 1.04 4.78 1.04 4.78 Stdev 0.797 13.9 0.805 13.7 0.805 13.7 p (t-test) 0.013 0.017 0.017 Min 0.00242 0.197 0.00242 0.197 0.00242 0.197 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 81 51 79 53 79 53 sCr only Median 0.950 1.35 0.950 1.35 0.950 1.35 Average 0.990 4.19 0.990 4.19 0.990 4.19 Stdev 0.779 12.5 0.779 12.5 0.779 12.5 p (t-test) 0.037 0.037 0.037 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 68 64 68 64 68 64 UO only Median 1.02 1.39 1.02 1.35 1.02 1.35 Average 1.09 5.46 1.10 5.26 1.10 5.18 Stdev 0.828 15.0 0.836 14.7 0.841 14.5 p (t-test) 0.0073 0.0098 0.011 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 87 44 85 46 84 47 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.66 0.64 0.65 0.66 0.63 0.65 0.66 0.63 SE 0.050 0.047 0.053 0.050 0.047 0.052 0.050 0.047 0.052 p Value 0.0020 7.5E−4 0.0087 0.0029 7.5E−4 0.012 0.0029 7.5E−4 0.011 nCohort Recovered 81 68 87 79 68 85 79 68 84 nCohort Non- 51 64 44 53 64 46 53 64 47 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Sensitivity 86% 84% 84% 87% 84% 85% 87% 84% 85% Specificity 32% 34% 30% 33% 34% 31% 33% 34% 31% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 59% 61% 61% 58% 61% 61% 58% 61% 62% Specificity 56% 60% 55% 56% 60% 55% 56% 60% 56% Cutoff Quartile 4 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Sensitivity 35% 34% 34% 34% 34% 33% 34% 34% 32% Specificity 81% 84% 79% 81% 84% 79% 81% 84% 79% OR Quartile 2 2.97 2.76 2.25 3.22 2.76 2.46 3.22 2.76 2.56 p Value 0.021 0.018 0.087 0.013 0.018 0.058 0.013 0.018 0.047 Lower limit of 95% CI 1.18 1.19 0.890 1.28 1.19 0.971 1.28 1.19 1.01 Upper limit of 95% CI 7.49 6.40 5.71 8.12 6.40 6.21 8.12 6.40 6.47 OR Quartile 3 1.79 2.37 1.95 1.77 2.37 1.92 1.77 2.37 2.05 p Value 0.11 0.016 0.076 0.11 0.016 0.079 0.11 0.016 0.054 Lower limit of 95% CI 0.879 1.18 0.933 0.876 1.18 0.927 0.876 1.18 0.987 Upper limit of 95% CI 3.63 4.76 4.09 3.58 4.76 3.99 3.58 4.76 4.24 OR Quartile 4 2.40 2.71 1.98 2.19 2.71 1.80 2.19 2.71 1.72 p Value 0.033 0.018 0.098 0.054 0.018 0.15 0.054 0.018 0.19 Lower limit of 95% CI 1.08 1.19 0.881 0.986 1.19 0.804 0.986 1.19 0.769 Upper limit of 95% CI 5.36 6.20 4.46 4.88 6.20 4.04 4.88 6.20 3.84

Example 23. Use of C—C Motif Chemokine 18 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 18 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 23.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.822 1.30 0.822 1.30 0.822 1.28 Average 0.944 3.40 0.944 3.40 0.955 3.34 Stdev 0.827 10.9 0.827 10.9 0.843 10.7 p (t-test) 0.13 0.13 0.15 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 46 86 46 86 44 88 sCr only Median 0.819 1.34 0.819 1.34 0.870 1.31 Average 0.934 3.71 0.934 3.71 0.952 3.63 Stdev 0.778 11.5 0.778 11.5 0.787 11.4 p (t-test) 0.077 0.077 0.090 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 55 76 55 76 53 78 UO only Median 1.02 1.42 0.974 1.43 1.00 1.36 Average 1.71 4.64 1.70 4.33 1.74 4.02 Stdev 6.16 13.3 6.33 12.5 6.48 12.0 p (t-test) 0.086 0.11 0.16 Min 0.00242 0.273 0.00242 0.273 0.00242 0.273 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 93 38 88 43 84 47 Recovery Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.66 0.67 0.66 0.66 0.67 0.69 0.65 0.66 0.66 SE 0.048 0.047 0.055 0.048 0.047 0.051 0.049 0.047 0.051 p Value 8.0E−4 2.0E−4 0.0031 8.0E−4 2.0E−4 2.0E−4 0.0020 8.6E−4 0.0019 nCohort 46 55 93 46 55 88 44 53 84 Recovered nCohort Non- 86 76 38 86 76 43 88 78 47 recovered Cutoff Quartile 2 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Sensitivity 81% 83% 87% 81% 83% 88% 81% 81% 87% Specificity 37% 36% 30% 37% 36% 32% 36% 34% 32% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 58% 61% 68% 58% 61% 70% 57% 59% 64% Specificity 65% 64% 57% 65% 64% 59% 64% 62% 57% Cutoff Quartile 4 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Sensitivity 29% 33% 34% 29% 33% 37% 28% 32% 34% Specificity 83% 85% 78% 83% 85% 81% 82% 85% 80% OR Quartile 2 2.56 2.77 2.84 2.56 2.77 3.55 2.39 2.16 3.24 p Value 0.022 0.014 0.049 0.022 0.014 0.016 0.035 0.059 0.018 Lower limit of 1.14 1.23 1.01 1.14 1.23 1.26 1.06 0.970 1.23 95% CI Upper limit of 5.76 6.23 8.04 5.76 6.23 9.98 5.37 4.81 8.55 95% CI OR Quartile 3 2.60 2.68 2.87 2.60 2.68 3.33 2.30 2.37 2.35 p Value 0.012 0.0069 0.0096 0.012 0.0069 0.0024 0.028 0.018 0.023 Lower limit of 1.24 1.31 1.29 1.24 1.31 1.53 1.09 1.16 1.13 95% CI Upper limit of 5.47 5.49 6.37 5.47 5.49 7.25 4.85 4.85 4.91 95% CI OR Quartile 4 1.95 2.88 1.90 1.95 2.88 2.47 1.79 2.65 2.03 p Value 0.14 0.020 0.13 0.14 0.020 0.029 0.20 0.032 0.084 Lower limit of 0.797 1.18 0.825 0.797 1.18 1.10 0.729 1.09 0.910 95% CI Upper limit of 4.76 7.01 4.37 4.76 7.01 5.59 4.37 6.46 4.55 95% CI

TABLE 23.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.940 1.33 0.940 1.33 0.960 1.32 Average 0.935 3.76 0.935 3.76 0.944 3.68 Stdev 0.763 11.6 0.763 11.6 0.774 11.5 p (t-test) 0.069 0.069 0.079 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 57 75 57 75 55 77 sCr only Median 0.905 1.36 0.905 1.36 0.950 1.35 Average 0.928 4.06 0.928 4.06 0.943 3.96 Stdev 0.731 12.1 0.731 12.1 0.738 12.0 p (t-test) 0.041 0.041 0.050 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 64 68 64 68 62 70 UO only Median 0.983 1.36 0.974 1.42 1.00 1.36 Average 1.71 4.72 1.69 4.60 1.75 4.10 Stdev 6.13 13.4 6.20 13.1 6.41 12.2 p (t-test) 0.080 0.086 0.15 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 94 37 92 39 86 45 Recovery Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.67 0.69 0.67 0.67 0.69 0.69 0.66 0.67 0.65 SE 0.046 0.046 0.055 0.046 0.046 0.053 0.047 0.046 0.052 p Value 2.0E−4 3.2E−5 0.0023 2.0E−4 3.2E−5 3.9E−4 5.4E−4 1.7E−4 0.0039 nCohort 57 64 94 57 64 92 55 62 86 Recovered nCohort Non- 75 68 37 75 68 39 77 70 45 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Sensitivity 84% 85% 89% 84% 85% 90% 83% 83% 89% Specificity 37% 36% 31% 37% 36% 32% 36% 34% 33% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 59% 60% 70% 59% 60% 72% 57% 59% 64% Specificity 61% 61% 57% 61% 61% 59% 60% 60% 57% Cutoff Quartile 4 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Sensitivity 33% 37% 32% 33% 37% 36% 32% 36% 31% Specificity 86% 88% 78% 86% 88% 79% 85% 87% 78% OR Quartile 2 3.06 3.25 3.68 3.06 3.25 4.03 2.81 2.48 3.86 p Value 0.0074 0.0061 0.023 0.0074 0.0061 0.015 0.012 0.029 0.010 Lower limit of 1.35 1.40 1.19 1.35 1.40 1.31 1.25 1.10 1.37 95% CI Upper limit of 6.95 7.56 11.4 6.95 7.56 12.4 6.33 5.59 10.9 95% CI OR Quartile 3 2.26 2.37 3.19 2.26 2.37 3.62 2.00 2.09 2.40 p Value 0.023 0.016 0.0053 0.023 0.016 0.0019 0.053 0.037 0.021 Lower limit of 1.12 1.18 1.41 1.12 1.18 1.61 0.990 1.04 1.14 95% CI Upper limit of 4.57 4.76 7.21 4.57 4.76 8.14 4.04 4.19 5.06 95% CI OR Quartile 4 3.06 4.07 1.67 3.06 4.07 2.15 2.82 3.75 1.59 p Value 0.014 0.0020 0.23 0.014 0.0020 0.069 0.022 0.0036 0.26 Lower limit of 1.26 1.67 0.719 1.26 1.67 0.942 1.16 1.54 0.708 95% CI Upper limit of 7.44 9.91 3.87 7.44 9.91 4.92 6.87 9.12 3.58 95% CI

TABLE 23.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.970 1.35 0.970 1.35 0.970 1.33 Average 0.991 4.05 0.991 4.05 0.987 3.96 Stdev 0.774 12.2 0.774 12.2 0.785 12.1 p (t-test) 0.047 0.047 0.053 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 65 67 65 67 63 69 sCr only Median 0.960 1.36 0.960 1.36 0.965 1.36 Average 0.948 4.18 0.948 4.18 0.955 4.13 Stdev 0.723 12.4 0.723 12.4 0.726 12.3 p (t-test) 0.035 0.035 0.039 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 67 65 67 65 66 66 UO only Median 1.02 1.36 1.02 1.36 1.02 1.35 Average 1.72 4.78 1.72 4.78 1.76 4.27 Stdev 6.10 13.6 6.10 13.6 6.30 12.7 p (t-test) 0.078 0.078 0.13 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 95 36 95 36 89 42 Recovery Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.65 0.68 0.64 0.65 0.68 0.64 0.65 0.67 0.63 SE 0.048 0.047 0.056 0.048 0.047 0.056 0.047 0.047 0.053 p Value 0.0014 1.1E−4 0.012 0.0014 1.1E−4 0.012 0.0014 2.5E−4 0.012 nCohort 65 67 95 65 67 95 63 66 89 Recovered nCohort Non- 67 65 36 67 65 36 69 66 42 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Sensitivity 84% 85% 86% 84% 85% 86% 84% 83% 88% Specificity 34% 34% 29% 34% 34% 29% 35% 33% 31% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 58% 60% 64% 58% 60% 64% 58% 59% 62% Specificity 58% 60% 55% 58% 60% 55% 59% 59% 55% Cutoff Quartile 4 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Sensitivity 34% 37% 33% 34% 37% 33% 33% 36% 31% Specificity 85% 87% 78% 85% 87% 78% 84% 86% 78% OR Quartile 2 2.60 2.88 2.59 2.60 2.88 2.59 2.83 2.50 3.40 p Value 0.023 0.014 0.073 0.023 0.014 0.073 0.014 0.030 0.021 Lower limit of 1.14 1.24 0.914 1.14 1.24 0.914 1.24 1.10 1.21 95% CI Upper limit of 5.95 6.67 7.35 5.95 6.67 7.35 6.47 5.71 9.57 95% CI OR Quartile 3 1.96 2.22 2.14 1.96 2.22 2.14 1.96 2.09 1.99 p Value 0.057 0.025 0.059 0.057 0.025 0.059 0.056 0.038 0.072 Lower limit of 0.981 1.11 0.970 0.981 1.11 0.970 0.982 1.04 0.940 95% CI Upper limit of 3.92 4.46 4.72 3.92 4.46 4.72 3.92 4.18 4.21 95% CI OR Quartile 4 2.87 3.77 1.76 2.87 3.77 1.76 2.65 3.62 1.55 p Value 0.014 0.0026 0.19 0.014 0.0026 0.19 0.023 0.0035 0.30 Lower limit of 1.24 1.59 0.756 1.24 1.59 0.756 1.14 1.53 0.680 95% CI Upper limit of 6.67 8.95 4.10 6.67 8.95 4.10 6.14 8.58 3.52 95% CI AUC 0.65 0.68 0.64 0.65 0.68 0.64 0.65 0.67 0.63 SE 0.048 0.047 0.056 0.048 0.047 0.056 0.047 0.047 0.053 p Value 0.0014 1.1E−4 0.012 0.0014 1.1E−4 0.012 0.0014 2.5E−4 0.012 nCohort 65 67 95 65 67 95 63 66 89 Recovered nCohort Non- 67 65 36 67 65 36 69 66 42 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569

TABLE 23.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.970 1.35 0.979 1.34 0.979 1.33 Average 1.84 3.31 1.86 3.27 1.88 3.20 Stdev 7.14 10.4 7.20 10.3 7.30 10.2 p (t-test) 0.34 0.36 0.39 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 69 63 68 64 66 66 sCr only Median 0.965 1.36 0.965 1.36 0.970 1.36 Average 1.79 3.39 1.79 3.39 1.81 3.34 Stdev 7.09 10.5 7.09 10.5 7.14 10.4 p (t-test) 0.30 0.30 0.32 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 70 62 70 62 69 63 UO only Median 1.02 1.36 1.04 1.36 1.02 1.35 Average 1.73 4.44 1.75 4.35 1.76 4.14 Stdev 6.23 13.0 6.26 12.8 6.37 12.4 p (t-test) 0.11 0.12 0.15 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 91 40 90 41 87 44 Recovery Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.65 0.67 0.64 0.64 0.67 0.63 0.64 0.66 0.63 SE 0.048 0.047 0.054 0.048 0.047 0.054 0.048 0.048 0.053 p Value 0.0024 4.1E−4 0.0096 0.0040 4.1E−4 0.016 0.0040 8.4E−4 0.012 nCohort 69 70 91 68 70 90 66 69 87 Recovered nCohort Non- 63 62 40 64 62 41 66 63 44 recovered Cutoff Quartile 2 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Sensitivity 83% 84% 85% 83% 84% 85% 83% 83% 86% Specificity 32% 33% 30% 32% 33% 30% 33% 32% 31% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 59% 60% 62% 58% 60% 61% 58% 59% 61% Specificity 58% 59% 55% 57% 59% 54% 58% 58% 55% Cutoff Quartile 4 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Sensitivity 35% 37% 35% 34% 37% 34% 33% 37% 32% Specificity 84% 86% 79% 84% 86% 79% 83% 86% 78% OR Quartile 2 2.21 2.54 2.39 2.30 2.54 2.50 2.50 2.21 2.85 p Value 0.059 0.029 0.081 0.047 0.029 0.066 0.030 0.059 0.035 Lower limit of 0.970 1.10 0.899 1.01 1.10 0.942 1.10 0.970 1.08 95% CI Upper limit of 5.05 5.90 6.35 5.26 5.90 6.64 5.71 5.05 7.54 95% CI OR Quartile 3 1.96 2.09 2.03 1.84 2.09 1.87 1.84 1.96 1.95 p Value 0.056 0.037 0.068 0.083 0.037 0.10 0.083 0.056 0.076 Lower limit of 0.982 1.04 0.949 0.924 1.04 0.880 0.923 0.982 0.933 95% CI Upper limit of 3.92 4.19 4.35 3.68 4.19 3.% 3.67 3.92 4.09 95% CI OR Quartile 4 2.83 3.54 2.04 2.71 3.54 1.94 2.50 3.39 1.67 p Value 0.014 0.0034 0.090 0.018 0.0034 0.11 0.030 0.0046 0.22 Lower limit of 1.24 1.52 0.896 1.19 1.52 0.853 1.10 1.46 0.741 95% CI Upper limit of 6.47 8.24 4.65 6.20 8.24 4.40 5.71 7.89 3.77 95% CI

TABLE 23.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 1.02 1.36 1.04 1.35 1.02 1.35 Average 1.73 4.22 1.75 4.13 1.75 4.07 Stdev 6.30 12.5 6.33 12.4 6.37 12.2 p (t-test) 0.13 0.14 0.15 Min 0.00242 0.197 0.00242 0.197 0.00242 0.197 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 89 43 88 44 87 45 sCr only Median 0.989 1.36 0.989 1.36 0.989 1.36 Average 1.78 3.68 1.78 3.68 1.78 3.68 Stdev 6.68 11.3 6.68 11.3 6.68 11.3 p (t-test) 0.23 0.23 0.23 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 79 53 79 53 79 53 UO only Median 1.05 1.35 1.05 1.35 1.05 1.34 Average 1.74 4.43 1.75 4.33 1.76 4.26 Stdev 6.23 13.0 6.26 12.8 6.30 12.7 p (t-test) 0.11 0.12 0.13 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 91 40 90 41 89 42 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.64 0.63 0.62 0.64 0.62 0.63 0.64 0.62 SE 0.053 0.050 0.054 0.053 0.050 0.054 0.052 0.050 0.054 p Value 0.011 0.0054 0.017 0.018 0.0054 0.027 0.016 0.0054 0.023 nCohort 89 79 91 88 79 90 87 79 89 Recovered nCohort 43 53 40 44 53 41 45 53 42 Non-recovered Cutoff 0.561 0.561 0.569 0.561 0.561 0.569 0.561 0.561 0.569 Quartile 2 Sensitivity 84% 81% 85% 84% 81% 85% 84% 81% 86% Specificity 29% 29% 30% 30% 29% 30% 30% 29% 30% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 60% 62% 60% 59% 62% 59% 60% 62% 60% Specificity 55% 58% 54% 55% 58% 53% 55% 58% 54% Cutoff 1.65 1.65 1.66 1.65 1.65 1.66 1.65 1.65 1.66 Quartile 4 Sensitivity 35% 36% 35% 34% 36% 34% 33% 36% 33% Specificity 80% 82% 79% 80% 82% 79% 79% 82% 79% OR Quartile 2 2.12 1.77 2.39 2.22 1.77 2.50 2.31 1.77 2.61 p Value 0.11 0.19 0.081 0.093 0.19 0.066 0.076 0.19 0.054 Lower limit 0.838 0.761 0.899 0.876 0.761 0.942 0.915 0.761 0.985 of 95% CI Upper limit 5.38 4.10 6.35 5.61 4.10 6.64 5.85 4.10 6.93 of 95% CI OR Quartile 3 1.87 2.30 1.75 1.73 2.30 1.61 1.85 2.30 1.72 p Value 0.097 0.022 0.15 0.14 0.022 0.21 0.10 0.022 0.15 Lower limit 0.894 1.13 0.823 0.833 1.13 0.765 0.889 1.13 0.818 of 95% CI Upper limit 3.93 4.69 3.72 3.61 4.69 3.40 3.83 4.69 3.62 of 95% CI OR Quartile 4 2.11 2.59 2.04 2.01 2.59 1.94 1.92 2.59 1.84 p Value 0.071 0.020 0.090 0.091 0.020 0.11 0.11 0.020 0.14 Lower limit 0.937 1.16 0.896 0.894 1.16 0.853 0.854 1.16 0.813 of 95% CI Upper limit 4.76 5.81 4.65 4.52 5.81 4.40 4.30 5.81 4.17 of 95% CI

Example 24. Use of C—C Motif Chemokine 18 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 18 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 24.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-Persistent Persistent Non-Persistent Persistent Non-Persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.989 1.36 0.978 1.54 0.989 1.54 Average 1.70 4.28 1.66 4.64 1.68 4.94 Stdev 6.30 12.5 6.16 13.1 6.04 13.8 p (t-test) 0.12 0.078 0.062 Min 0.00242 0.233 0.00242 0.233 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 89 43 93 39 97 35 sCr only Median 0.978 1.39 0.983 1.59 1.00 1.54 Average 1.69 4.37 1.65 4.81 1.69 4.90 Stdev 6.30 12.6 6.13 13.4 6.07 13.8 p (t-test) 0.11 0.066 0.066 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 89 42 94 37 96 35 UO only Median 1.06 1.58 1.06 1.74 1.06 1.74 Average 1.65 8.26 1.64 9.16 1.64 9.16 Stdev 5.60 18.9 5.55 19.9 5.55 19.9 p (t-test) 0.0030 0.0013 0.0013 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 113 18 115 16 115 16 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.68 0.71 0.70 0.70 0.73 0.68 0.67 0.73 SE 0.052 0.052 0.072 0.053 0.054 0.075 0.055 0.056 0.075 p Value 0.0013 4.8E−4 0.0046 2.2E−4 2.6E−4 0.0018 8.4E−4 0.0026 0.0018 nCohort 89 89 113 93 94 115 97 96 115 Non-persistent nCohort 43 42 18 39 37 16 35 35 16 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 88% 88% 94% 87% 86% 94% 89% 86% 94% Specificity 31% 31% 28% 30% 30% 28% 30% 29% 28% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 65% 69% 72% 72% 70% 75% 71% 69% 75% Specificity 57% 58% 53% 59% 57% 53% 58% 56% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 35% 36% 44% 38% 41% 50% 37% 37% 50% Specificity 80% 80% 78% 81% 81% 78% 79% 79% 78% OR Quartile 2 3.49 3.40 6.72 2.93 2.72 5.78 3.31 2.47 5.78 p Value 0.018 0.021 0.070 0.042 0.060 0.096 0.038 0.090 0.096 Lower limit 1.24 1.21 0.858 1.04 0.959 0.733 1.07 0.870 0.733 of 95% CI Upper limit 9.81 9.57 52.6 8.27 7.69 45.6 10.2 7.02 45.6 of 95% CI OR Quartile 3 2.51 3.14 2.94 3.68 3.19 3.39 3.41 2.81 3.39 p Value 0.017 0.0040 0.053 0.0016 0.0053 0.044 0.0040 0.014 0.044 Lower limit 1.18 1.44 0.984 1.64 1.41 1.03 1.48 1.24 1.03 of 95% CI Upper limit 5.33 6.83 8.80 8.29 7.21 11.1 7.88 6.37 11.1 of 95% CI OR Quartile 4 2.11 2.19 2.82 2.60 2.88 3.60 2.28 2.25 3.60 p Value 0.071 0.060 0.049 0.023 0.013 0.020 0.056 0.060 0.020 Lower limit 0.937 0.969 1.00 1.14 1.25 1.23 0.978 0.965 1.23 of 95% CI Upper limit 4.76 4.96 7.89 5.94 6.63 10.6 5.29 5.22 10.6 of 95% CI

TABLE 24.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent'’ and “non-persistent″ cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-Persistent Persistent Non-Persistent Persistent Non-Persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.970 1.36 0.960 1.54 0.970 1.54 Average 1.73 3.75 1.67 4.12 1.68 4.32 Stdev 6.68 11.3 6.45 12.0 6.30 12.5 p (t-test) 0.20 0.13 0.11 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 79 53 85 47 89 43 sCr only Median 0.965 1.36 0.965 1.43 0.978 1.39 Average 1.72 3.84 1.68 4.09 1.71 4.20 Stdev 6.64 11.5 6.48 12.0 6.37 12.4 p (t-test) 0.18 0.14 0.13 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 80 51 84 47 87 44 UO only Median 1.05 1.48 1.05 1.58 1.05 1.58 Average 1.67 6.52 1.66 7.02 1.66 7.02 Stdev 5.75 16.5 5.70 17.2 5.70 17.2 p (t-test) 0.015 0.0093 0.0093 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 107 24 109 22 109 22 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.68 0.68 0.71 0.70 0.69 0.70 0.67 0.69 SE 0.049 0.049 0.065 0.049 0.050 0.067 0.051 0.052 0.067 p Value 3.3E−4 2.6E−4 0.0068 2.0E−5 8.2E−5 0.0039 7.5E−5 9.1E−4 0.0039 nCohort 79 80 107 85 84 109 89 87 109 Non-persistent nCohort 53 51 24 47 47 22 43 44 22 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 87% 86% 92% 87% 87% 91% 88% 86% 91% Specificity 33% 32% 29% 32% 32% 28% 31% 31% 28% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 62% 65% 67% 68% 66% 68% 70% 66% 68% Specificity 58% 59% 53% 60% 58% 53% 60% 57% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 38% 37% 42% 40% 38% 45% 40% 36% 45% Specificity 84% 82% 79% 84% 82% 79% 82% 80% 79% OR Quartile 2 3.22 3.03 4.49 3.18 3.24 3.97 3.49 2.85 3.97 p Value 0.013 0.019 0.051 0.019 0.018 0.074 0.018 0.035 0.074 Lower limit 1.28 1.20 0.995 1.20 1.23 0.876 1.24 1.08 0.876 of 95% CI Upper limit 8.12 7.63 20.2 8.40 8.55 18.0 9.81 7.54 18.0 of 95% CI OR Quartile 3 2.30 2.61 2.28 3.20 2.71 2.44 3.40 2.61 2.44 p Value 0.022 0.0096 0.082 0.0024 0.0085 0.073 0.0020 0.013 0.073 Lower limit 1.13 1.26 0.900 1.51 1.29 0.921 1.56 1.23 0.921 of 95% CI Upper limit 4.69 5.40 5.78 6.78 5.70 6.45 7.38 5.56 6.45 of 95% CI OR Quartile 4 3.08 2.80 2.61 3.44 2.86 3.12 2.98 2.35 3.12 p Value 0.0068 0.013 0.044 0.0030 0.011 0.020 0.0087 0.039 0.020 Lower limit 1.36 1.25 1.03 1.52 1.27 1.20 1.32 1.05 1.20 of 95% CI Upper limit 6.94 6.29 6.63 7.79 6.42 8.11 6.75 5.30 8.11 of 95% CI

TABLE 24.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-Persistent Persistent Non-Persistent Persistent Non-Persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.965 1.35 0.960 1.36 0.965 1.39 Average 1.79 3.50 1.73 3.75 1.72 3.88 Stdev 6.90 10.8 6.68 11.3 6.56 11.6 p (t-test) 0.27 0.20 0.17 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 74 58 79 53 82 50 sCr only Median 0.960 1.36 0.960 1.39 0.970 1.39 Average 1.75 3.68 1.71 3.90 1.72 3.98 Stdev 6.77 11.2 6.60 11.6 6.52 11.9 p (t-test) 0.22 0.17 0.16 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 77 54 81 50 83 48 UO only Median 1.05 1.36 1.05 1.45 1.05 1.45 Average 1.69 5.61 1.67 6.16 1.67 6.16 Stdev 5.89 15.1 5.80 15.9 5.80 15.9 p (t-test) 0.035 0.020 0.020 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 102 29 105 26 105 26 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.67 0.64 0.68 0.69 0.67 0.68 0.67 0.67 SE 0.048 0.049 0.061 0.049 0.049 0.063 0.049 0.050 0.063 p Value 0.0011 4.5E−4 0.018 2.3E−4 1.6E−4 0.0058 2.3E−4 5.2E−4 0.0058 nCohort 74 77 102 79 81 105 82 83 105 Non-persistent nCohort 58 54 29 53 50 26 50 48 26 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 84% 85% 86% 85% 86% 88% 86% 85% 88% Specificity 32% 32% 28% 32% 32% 29% 32% 31% 29% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 60% 63% 62% 64% 64% 65% 66% 65% 65% Specificity 58% 58% 53% 59% 58% 53% 60% 58% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 34% 35% 38% 36% 36% 42% 36% 35% 42% Specificity 82% 82% 78% 82% 81% 79% 82% 81% 79% OR Quartile 2 2.61 2.76 2.48 2.60 2.90 3.07 2.85 2.67 3.07 p Value 0.029 0.025 0.12 0.035 0.024 0.085 0.026 0.038 0.085 Lower limit 1.10 1.14 0.794 1.07 1.15 0.857 1.13 1.06 0.857 of 95% CI Upper limit 6.18 6.73 7.76 6.34 7.32 11.0 7.19 6.74 11.0 of 95% CI OR Quartile 3 2.11 2.39 1.84 2.63 2.46 2.16 2.88 2.50 2.16 p Value 0.036 0.017 0.16 0.0084 0.015 0.092 0.0046 0.014 0.092 Lower limit 1.05 1.17 0.791 1.28 1.19 0.883 1.38 1.20 0.883 of 95% CI Upper limit 4.25 4.88 4.29 5.39 5.08 5.28 6.00 5.21 5.28 of 95% CI OR Quartile 4 2.47 2.44 2.22 2.59 2.47 2.77 2.51 2.30 2.77 p Value 0.028 0.030 0.077 0.020 0.027 0.028 0.025 0.043 0.028 Lower limit 1.10 1.09 0.916 1.16 1.11 1.11 1.12 1.03 1.11 of 95% CI Upper limit 5.54 5.46 5.39 5.81 5.53 6.87 5.62 5.13 6.87 of 95% CI

TABLE 24.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.960 1.36 0.950 1.39 0.965 1.39 Average 1.79 3.47 1.73 3.71 1.74 3.77 Stdev 6.95 10.7 6.73 11.2 6.64 11.4 p (t-test) 0.28 0.21 0.20 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 73 59 78 54 80 52 sCr only Median 0.960 1.36 0.960 1.39 0.970 1.39 Average 1.77 3.59 1.73 3.79 1.74 3.86 Stdev 6.86 11.0 6.68 11.4 6.60 11.6 p (t-test) 0.25 0.19 0.18 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 75 56 79 52 81 50 UO only Median 1.05 1.36 1.05 1.45 1.05 1.45 Average 1.70 5.11 1.67 5.54 1.67 5.54 Stdev 6.01 14.2 5.92 14.8 5.92 14.8 p (t-test) 0.056 0.036 0.036 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 98 33 101 30 101 30 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.67 0.67 0.65 0.69 0.68 0.67 0.67 0.67 0.67 SE 0.048 0.048 0.058 0.048 0.049 0.059 0.049 0.050 0.059 p Value 5.6E−4 4.5E−4 0.011 1.0E−4 1.7E−4 0.0037 3.5E−4 5.5E−4 0.0037 nCohort Non- 73 75 98 78 79 101 80 81 101 persistent nCohort 59 56 33 54 52 30 52 50 30 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 85% 84% 88% 85% 85% 90% 85% 84% 90% Specificity 33% 32% 30% 32% 32% 30% 31% 31% 30% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 61% 62% 61% 65% 63% 63% 65% 64% 63% Specificity 59% 59% 53% 60% 58% 53% 60% 58% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 36% 36% 39% 37% 37% 43% 37% 36% 43% Specificity 84% 83% 80% 83% 82% 80% 82% 81% 80% OR 2.72 2.46 3.05 2.71 2.55 3.80 2.50 2.34 3.80 Quartile 2 p Value 0.023 0.041 0.054 0.028 0.040 0.039 0.043 0.061 0.039 Lower limit 1.15 1.04 0.983 1.12 1.05 1.07 1.03 0.961 1.07 of 95% CI Upper limit 6.44 5.82 9.45 6.60 6.20 13.5 6.08 5.71 13.5 of 95% CI OR Quartile 3 2.24 2.37 1.74 2.79 2.42 1.98 2.83 2.46 1.98 p Value 0.024 0.017 0.18 0.0051 0.016 0.11 0.0049 0.015 0.11 Lower limit 1.11 1.16 0.779 1.36 1.18 0.857 1.37 1.19 0.857 of 95% CI Upper limit 4.52 4.81 3.88 5.73 4.97 4.59 5.85 5.08 4.59 of 95% CI OR Quartile 4 2.81 2.65 2.54 2.94 2.67 3.10 2.71 2.47 3.10 p Value 0.013 0.018 0.033 0.0092 0.017 0.011 0.015 0.027 0.011 Lower limit 1.24 1.18 1.08 1.31 1.19 1.29 1.21 1.11 1.29 of 95% CI Upper limit 6.36 5.96 5.95 6.63 5.99 7.41 6.08 5.53 7.41 of 95% CI

TABLE 24.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.960 1.36 0.950 1.39 0.965 1.39 Average 1.79 3.47 1.73 3.71 1.74 3.77 Stdev 6.95 10.7 6.73 11.2 6.64 11.4 p (t-test) 0.28 0.21 0.20 Min 0.0156 0.00242 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 73 59 78 54 80 52 sCr only Median 0.965 1.36 0.965 1.36 0.979 1.36 Average 1.79 3.53 1.75 3.72 1.76 3.78 Stdev 6.90 10.9 6.72 11.3 6.64 11.5 p (t-test) 0.27 0.21 0.20 Min 0.0264 0.00242 0.00242 0.0156 0.00242 0.0156 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 74 57 78 53 80 51 UO only Median 1.05 1.36 1.05 1.36 1.05 1.36 Average 1.71 4.88 1.70 5.12 1.70 5.12 Stdev 6.07 13.8 6.01 14.2 6.01 14.2 p (t-test) 0.070 0.055 0.055 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 96 35 98 33 98 33 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.67 0.65 0.64 0.69 0.67 0.65 0.67 0.65 0.65 SE 0.048 0.049 0.057 0.048 0.049 0.058 0.049 0.050 0.058 p Value 5.6E−4 0.0016 0.013 1.0E−4 7.0E−4 0.0093 3.5E−4 0.0020 0.0093 nCohort Non- 73 74 96 78 78 98 80 80 98 persistent nCohort 59 57 35 54 53 33 52 51 33 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 85% 82% 89% 85% 83% 88% 85% 82% 88% Specificity 33% 31% 30% 32% 31% 30% 31% 30% 30% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 61% 61% 60% 65% 62% 61% 65% 63% 61% Specificity 59% 58% 53% 60% 58% 53% 60% 57% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 36% 35% 37% 37% 36% 39% 37% 35% 39% Specificity 84% 82% 79% 83% 82% 80% 82% 81% 80% OR Quartile 2 2.72 2.12 3.35 2.71 2.17 3.05 2.50 2.00 3.05 p Value 0.023 0.080 0.036 0.028 0.078 0.054 0.043 0.12 0.054 Lower limit 1.15 0.914 1.08 1.12 0.916 0.983 1.03 0.843 0.983 of 95% CI Upper limit 6.44 4.92 10.4 6.60 5.15 9.45 6.08 4.75 9.45 of 95% CI OR Quartile 3 2.24 2.21 1.70 2.79 2.25 1.74 2.83 2.28 1.74 p Value 0.024 0.028 0.19 0.0051 0.026 0.18 0.0049 0.025 0.18 Lower limit 1.11 1.09 0.775 1.36 1.10 0.779 1.37 1.11 0.779 of 95% CI Upper limit 4.52 4.47 3.73 5.73 4.60 3.88 5.85 4.68 3.88 of 95% CI OR Quartile 4 2.81 2.54 2.25 2.94 2.55 2.54 2.71 2.36 2.54 p Value 0.013 0.024 0.060 0.0092 0.023 0.033 0.015 0.036 0.033 Lower limit 1.24 1.13 0.965 1.31 1.14 1.08 1.21 1.06 1.08 of 95% CI Upper limit 6.36 5.70 5.22 6.63 5.72 5.95 6.08 5.28 5.95 of 95% CI

Example 25. Use of C—C Motif Chemokine 18 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 18 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 25.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.845 1.33 0.979 1.34 0.989 1.36 Average 0.977 3.63 1.02 4.25 1.02 4.55 Stdev 0.836 11.4 0.798 12.7 0.777 13.2 p (t-test) 0.091 0.036 0.023 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 54 78 70 62 75 57 sCr only Median 0.870 1.31 0.970 1.35 0.989 1.44 Average 0.984 3.75 1.000 4.38 1.00 4.75 Stdev 0.819 11.7 0.777 12.9 0.752 13.6 p (t-test) 0.077 0.030 0.017 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 57 74 71 60 77 54 UO only Median 0.983 1.59 1.02 1.59 1.04 1.61 Average 1.64 5.80 1.64 6.89 1.63 7.41 Stdev 5.89 15.1 5.73 16.8 5.67 17.5 p (t-test) 0.025 0.0095 0.0058 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 102 29 108 23 110 21 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.66 0.65 0.73 0.64 0.66 0.74 0.65 0.66 0.74 SE 0.047 0.048 0.057 0.048 0.048 0.063 0.049 0.049 0.066 p Value 0.0011 0.0017 5.0E−5 0.0038 0.0012 1.8E−4 0.0017 8.2E−4 2.7E−4 nCohort Non- 54 57 102 70 71 108 75 77 110 persistent nCohort Persistent 78 74 29 62 60 23 57 54 21 Cutoff Quartile 2 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Sensitivity 83% 82% 97% 85% 85% 96% 86% 85% 95% Specificity 37% 35% 31% 34% 34% 30% 33% 32% 29% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 59% 59% 79% 58% 62% 78% 60% 61% 76% Specificity 63% 61% 58% 57% 59% 56% 57% 57% 55% Cutoff Quartile 4 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Sensitivity 29% 31% 38% 32% 33% 43% 35% 37% 48% Specificity 81% 82% 78% 81% 82% 79% 83% 83% 79% OR Quartile 2 2.94 2.54 12.8 3.07 2.89 9.26 3.06 2.76 8.21 p Value 0.0092 0.024 0.014 0.011 0.016 0.033 0.014 0.025 0.044 Lower limit of 1.31 1.13 1.67 1.30 1.22 1.20 1.26 1.14 1.06 95% CI Upper limit of 6.63 5.70 98.3 7.27 6.86 71.7 7.44 6.73 63.7 95% CI OR Quartile 3 2.44 2.33 5.26 1.85 2.33 4.50 1.99 2.10 3.84 p Value 0.014 0.019 9.1E−4 0.082 0.018 0.0055 0.054 0.041 0.014 Lower limit of 1.20 1.15 1.97 0.924 1.15 1.56 0.987 1.03 1.31 95% CI Upper limit of 4.99 4.73 14.0 3.69 4.71 13.0 4.00 4.26 11.2 95% CI OR Quartile 4 1.84 2.12 2.22 2.09 2.23 2.84 2.58 2.90 3.44 p Value 0.16 0.080 0.077 0.073 0.051 0.030 0.022 0.010 0.013 Lower limit of 0.793 0.914 0.916 0.934 0.996 1.11 1.15 1.28 1.30 95% CI Upper limit of 4.27 4.92 5.39 4.67 5.00 7.31 5.79 6.53 9.09 95% CI

TABLE 25.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.774 1.33 0.965 1.31 0.965 1.35 Average 0.895 3.51 0.997 4.00 0.988 4.19 Stdev 0.778 11.0 0.784 12.2 0.766 12.5 p (t-test) 0.10 0.051 0.037 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 49 83 64 68 68 64 sCr only Median 0.797 1.35 0.950 1.35 0.960 1.36 Average 0.903 3.70 0.953 4.16 0.952 4.32 Stdev 0.750 11.5 0.734 12.4 0.721 12.7 p (t-test) 0.076 0.038 0.030 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 54 77 66 65 69 62 UO only Median 0.974 1.42 1.00 1.43 1.02 1.54 Average 1.69 4.77 1.68 5.39 1.68 5.66 Stdev 6.13 13.4 5.95 14.6 5.89 15.1 p (t-test) 0.073 0.042 0.032 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 94 37 100 31 102 29 Persistence Period Duration (hr) 24 48 72 sCr or UO sCr only UO only sCr or UO sCr only UO only sCr or UO sCr only UO only AUC 0.69 0.69 0.69 0.65 0.67 0.68 0.66 0.68 0.67 SE 0.046 0.046 0.054 0.048 0.047 0.058 0.047 0.047 0.060 p Value 4.11E−5 4.7E−5 5.7E−4 0.0023 3.2E−4 0.0024 8.2E−4 1.8E−4 0.0039 nCohort Non- 49 54 94 64 66 100 68 69 102 persistent nCohort Persistent 83 77 37 68 65 31 64 62 29 Cutoff Quartile 2 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Sensitivity 83% 83% 92% 84% 85% 90% 84% 85% 90% Specificity 39% 37% 32% 34% 35% 30% 34% 35% 29% Cutoff Quartile 3 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Sensitivity 60% 61% 73% 57% 60% 71% 59% 60% 69% Specificity 67% 65% 59% 58% 59% 56% 59% 58% 55% Cutoff Quartile 4 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Sensitivity 31% 34% 32% 32% 34% 35% 34% 35% 38% Specificity 86% 87% 78% 83% 83% 78% 84% 84% 78% OR Quartile 2 3.12 2.90 5.31 2.71 2.94 4.00 2.76 3.14 3.61 p Value 0.0060 0.010 0.0092 0.018 0.012 0.032 0.018 0.0093 0.047 Lower limit of 1.38 1.28 1.51 1.19 1.27 1.13 1.19 1.33 1.02 95% CI Upper limit of 7.03 6.53 18.7 6.20 6.83 14.2 6.40 7.44 12.8 95% CI OR Quartile 3 3.12 2.89 3.81 1.84 2.17 3.11 2.09 2.04 2.71 p Value 0.0026 0.0040 0.0017 0.083 0.030 0.011 0.038 0.045 0.026 Lower limit of 1.49 1.40 1.65 0.924 1.08 1.30 1.04 1.02 1.12 95% CI Upper limit of 6.56 5.94 8.76 3.68 4.35 7.43 4.18 4.10 6.51 95% CI OR Quartile 4 2.74 3.42 1.67 2.30 2.56 1.95 2.71 2.90 p Value 0.033 0.0090 0.23 0.047 0.026 0.13 0.018 0.012 0.077 Lower limit of 1.09 1.36 0.719 1.01 1.12 0.813 1.19 1.27 0.916 95% CI Upper limit of 6.90 8.62 3.87 5.26 5.85 4.68 6.20 6.64 5.39 95% CI

TABLE 25.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.774 1.33 0.965 1.31 0.979 1.34 Average 0.895 3.51 0.997 4.00 1.00 4.08 Stdev 0.778 11.0 0.784 12.2 0.773 12.3 p (t-test) 0.10 0.051 0.045 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 49 83 64 68 66 66 sCr only Median 0.819 1.34 0.960 1.34 0.970 1.35 Average 0.911 3.66 0.960 4.11 0.967 4.20 Stdev 0.755 11.4 0.737 12.3 0.727 12.5 p (t-test) 0.082 0.042 0.037 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 53 78 65 66 67 64 UO only Median 1.02 1.35 1.05 1.35 1.06 1.34 Average 1.75 4.29 1.74 4.73 1.73 4.93 Stdev 6.30 12.7 6.10 13.6 6.04 14.0 p (t-test) 0.13 0.085 0.071 Min 0.00242 0.310 0.00242 0.310 0.00242 0.310 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 89 42 95 36 97 34 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.68 0.64 0.65 0.66 0.62 0.64 0.66 0.62 SE 0.046 0.046 0.053 0.048 0.047 0.057 0.048 0.048 0.058 p Value 4.1E−5 1.1E−4 0.0096 0.0023 6.6E−4 0.031 0.0028 8.3E−4 0.046 nCohort 49 53 89 64 65 95 66 67 97 Non-persistent nCohort 83 78 42 68 66 36 66 64 34 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 83% 82% 88% 84% 83% 86% 83% 83% 85% Specificity 39% 36% 31% 34% 34% 29% 33% 33% 29% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 60% 60% 64% 57% 59% 61% 58% 58% 59% Specificity 67% 64% 56% 58% 58% 54% 58% 57% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 31% 33% 29% 32% 33% 31% 33% 34% 32% Specificity 86% 87% 76% 83% 83% 77% 83% 84% 77% OR Quartile 2 3.12 2.55 3.40 2.71 2.56 2.59 2.50 2.36 2.35 p Value 0.0060 0.023 0.021 0.018 0.026 0.073 0.030 0.042 0.11 Lower limit 1.38 1.14 1.21 1.19 1.12 0.914 1.10 1.03 0.827 of 95% CI Upper limit 7.03 5.72 9.57 6.20 5.85 7.35 5.71 5.38 6.70 of 95% CI OR Quartile 3 3.12 2.71 2.31 1.84 2.03 1.82 1.84 1.80 1.58 p Value 0.0026 0.0067 0.030 0.083 0.046 0.13 0.083 0.098 0.25 Lower limit 1.49 1.32 1.08 0.924 1.01 0.833 0.923 0.898 0.718 of 95% CI Upper limit 6.56 5.58 4.92 3.68 4.08 3.98 3.67 3.59 3.49 of 95% CI OR Quartile 4 2.74 3.29 1.30 2.30 2.45 1.46 2.50 2.67 1.63 p Value 0.033 0.012 0.54 0.047 0.033 0.39 0.030 0.020 0.27 Lower limit 1.09 1.30 0.565 1.01 1.07 0.621 1.10 1.17 0.689 of 95% CI Upper limit 6.90 8.28 2.97 5.26 5.61 3.43 5.71 6.10 3.86 of 95% CI

TABLE 25.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.774 1.33 0.960 1.33 0.970 1.35 Average 0.895 3.51 0.985 3.96 0.991 4.05 Stdev 0.778 11.0 0.785 12.1 0.774 12.2 p (t-test) 0.10 0.053 0.047 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 49 83 63 69 65 67 sCr only Median 0.819 1.34 0.950 1.35 0.965 1.36 Average 0.911 3.66 0.949 4.07 0.955 4.16 Stdev 0.755 11.4 0.736 12.2 0.726 12.4 p (t-test) 0.082 0.044 0.038 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 53 78 64 67 66 65 UO only Median 1.00 1.36 1.05 1.35 1.06 1.34 Average 1.75 4.11 1.76 4.39 1.75 4.55 Stdev 6.41 12.2 6.23 13.0 6.16 13.3 p (t-test) 0.15 0.12 0.10 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 86 45 91 40 93 38 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.68 0.64 0.65 0.67 0.62 0.65 0.67 0.61 SE 0.046 0.046 0.052 0.047 0.047 0.055 0.048 0.047 0.056 p Value 4.1E−5 4.4E−4 0.0059 0.0012 2.9E−4 0.0032 0.0014 3.8E−4 0.007 nCohort 49 53 86 63 64 91 65 66 93 Non-persistent nCohort 83 78 45 69 67 40 67 65 38 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 83% 82% 87% 84% 84% 85% 84% 83% 84% Specificity 39% 36% 31% 35% 34% 30% 34% 33% 29% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 60% 60% 64% 58% 60% 60% 58% 58% 58% Specificity 67% 64% 57% 59% 59% 54% 58% 58% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 31% 33% 31% 33% 34% 32% 34% 35% 34% Specificity 86% 87% 78% 84% 84% 78% 85% 85% 78% OR Quartile 2 3.12 2.55 2.97 2.83 2.67 2.39 2.60 2.45 2.18 p Value 0.0060 0.023 0.028 0.014 0.020 0.081 0.023 0.033 0.12 Lower limit 1.38 1.14 1.12 1.24 1.17 0.899 1.14 1.07 0.819 of 95% CI Upper limit 7.03 5.72 7.87 6.47 6.10 6.35 5.95 5.61 5.81 of 95% CI OR Quartile 3 3.12 2.71 2.40 1.96 2.17 1.75 1.96 1.91 1.53 p Value 0.0026 0.0067 0.021 0.056 0.030 0.15 0.057 0.068 0.27 Lower limit 1.49 1.32 1.14 0.982 1.08 0.823 0.981 0.954 0.715 of 95% CI Upper limit 6.56 5.58 5.06 3.92 4.35 3.72 3.92 3.82 3.28 of 95% CI OR Quartile 4 2.74 3.29 1.59 2.65 2.82 1.71 2.87 3.07 1.90 p Value 0.033 0.012 0.26 0.023 0.016 0.20 0.014 0.0092 0.13 Lower limit 1.09 1.30 0.708 1.14 1.22 0.748 1.24 1.32 0.825 of 95% CI Upper limit 6.90 8.28 3.58 6.14 6.55 3.91 6.67 7.13 4.37 of 95% CI

TABLE 25.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.774 1.33 0.950 1.33 0.965 1.34 Average 0.895 3.51 0.980 3.93 0.986 4.01 Stdev 0.778 11.0 0.791 12.0 0.779 12.2 p (t-test) 0.10 0.056 0.049 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 49 83 62 70 64 68 sCr only Median 0.819 1.34 0.950 1.35 0.965 1.36 Average 0.911 3.66 0.949 4.07 0.955 4.16 Stdev 0.755 11.4 0.736 12.2 0.726 12.4 p (t-test) 0.082 0.044 0.038 Min 0.0264 0.00242 0.0264 0.00242 0.0264 0.00242 Max 4.23 60.0 4.23 60.0 4.23 60.0 n (Patient) 53 78 64 67 66 65 UO only Median 1.00 1.36 1.04 1.35 1.05 1.33 Average 1.77 3.98 1.78 4.17 1.77 4.30 Stdev 6.48 12.0 6.33 12.5 6.26 12.8 p (t-test) 0.17 0.15 0.13 Min 0.00242 0.233 0.00242 0.233 0.00242 0.233 Max 60.0 60.0 60.0 60.0 60.0 60.0 n (Patient) 84 47 88 43 90 41 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.68 0.64 0.66 0.67 0.62 0.65 0.67 0.61 SE 0.046 0.046 0.052 0.047 0.047 0.054 0.047 0.047 0.054 p Value 4.1E−5 1.1E−4 0.0064 8.8E−4 2.9E−4 0.030 0.0011 3.8E−4 0.045 nCohort 49 53 84 62 64 88 64 66 90 Non-persistent nCohort 83 78 47 70 67 43 68 65 41 Persistent Cutoff 0.561 0.557 0.569 0.561 0.557 0.569 0.561 0.557 0.569 Quartile 2 Sensitivity 83% 82% 87% 84% 84% 86% 84% 83% 85% Specificity 39% 36% 32% 35% 34% 31% 34% 33% 30% Cutoff 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 1.07 Quartile 3 Sensitivity 60% 60% 64% 59% 60% 60% 59% 58% 59% Specificity 67% 64% 57% 60% 59% 55% 59% 58% 53% Cutoff 1.65 1.64 1.66 1.65 1.64 1.66 1.65 1.64 1.66 Quartile 4 Sensitivity 31% 33% 30% 33% 34% 30% 34% 35% 32% Specificity 86% 87% 77% 84% 84% 77% 84% 85% 78% OR Quartile 2 3.12 2.55 3.24 2.95 2.67 2.73 2.71 2.45 2.50 p Value 0.0060 0.023 0.018 0.010 0.020 0.043 0.018 0.033 0.066 Lower limit 1.38 1.14 1.23 1.29 1.17 1.03 1.19 1.07 0.942 of 95% CI Upper limit 7.03 5.72 8.55 6.75 6.10 7.23 6.20 5.61 6.64 of 95% CI OR Quartile 3 3.12 2.71 2.35 2.09 2.17 1.84 2.09 1.91 1.61 p Value 0.0026 0.0067 0.023 0.037 0.030 0.11 0.038 0.068 0.21 Lower limit 1.49 1.32 1.13 1.04 1.08 0.874 1.04 0.954 0.765 of 95% CI Upper limit 6.56 5.58 4.91 4.19 4.35 3.85 4.18 3.82 3.40 of 95% CI OR Quartile 4 2.74 3.29 1.45 2.54 2.82 1.47 2.76 3.07 1.62 p Value 0.033 0.012 0.37 0.029 0.016 0.35 0.018 0.0092 0.25 Lower limit 1.09 1.30 0.647 1.10 1.22 0.649 1.19 1.32 0.713 of 95% CI Upper limit 6.90 8.28 3.25 5.90 6.55 3.34 6.40 7.13 3.71 of 95% CI

Example 26. Use of C—C Motif Chemokine 23 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 23 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 26.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00282 0.0144 0.00258 0.0130 0.00282 0.0123 Average 0.00875 0.134 0.00707 0.129 0.00774 0.127 Stdev 0.0140 0.372 0.00989 0.366 0.0103 0.363 p (t-test) 0.012 0.025 0.037 Min 7.19E−7 3.28E−9 7.19E−7 3.28E−9 7.19E−7 3.28E−9 Max 0.0790 2.47 0.0380 2.47 0.0380 2.47 n (Patient) 57 260 46 271 41 276 sCr only Median 0.00278 0.0172 0.00278 0.0168 0.00282 0.0162 Average 0.00748 0.144 0.00760 0.143 0.00783 0.141 Stdev 0.0121 0.386 0.0123 0.384 0.0125 0.382 p (t-test) 0.0022 0.0027 0.0038 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 0.0790 2.47 0.0790 2.47 0.0790 2.47 n (Patient) 76 240 74 242 70 246 UO only Median 0.00780 0.0134 0.00783 0.0129 0.00764 0.0134 Average 0.0920 0.144 0.0859 0.144 0.0865 0.137 Stdev 0.347 0.331 0.338 0.345 0.350 0.332 p (t-test) 0.19 0.14 0.19 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 6.85E−7 3.28E−9 Max 2.47 1.82 2.47 1.92 2.47 1.92 n (Patient) 194 121 173 142 156 159 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.73 0.57 0.71 0.73 0.57 0.69 0.72 0.57 SE 0.035 0.030 0.033 0.037 0.030 0.033 0.040 0.031 0.032 p Value 9.9E−9 5.1E−15 0.036 1.9E−8 5.0E−14 0.034 2.0E−6 1.1E−12 0.020 nCohort 57 76 194 46 74 173 41 70 156 Recovered nCohort 260 240 121 271 242 142 276 246 159 Non-recovered Cutoff 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Quartile 2 Sensitivity 78% 80% 78%  78% 80% 78%  77% 80% 78% Specificity 40% 42% 27%  43% 42% 28%  39% 41% 28% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 56% 59% 55%  55% 59% 54%  54% 58% 54% Specificity 75% 79% 53%  76% 78% 53%  73% 77% 54% Cutoff 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Quartile 4 Sensitivity 30% 32% 36%  30% 32% 35%  29% 32% 33% Specificity 98% 99% 82% 100% 99% 83% 100% 99% 83% OR Quartile 2 2.46 2.99 1.27 2.76 2.91 1.37 2.16 2.77 1.39 p Value 0.0035 1.1E−4 0.37 0.0022 1.8E−4 0.23 0.028 4.3E−4 0.21 Lower limit 1.34 1.71 0.748 1.44 1.67 0.818 1.09 1.57 0.834 of 95% CI Upper limit 4.52 5.21 2.17 5.30 5.10 2.31 4.30 4.89 2.32 of 95% CI OR Quartile 3 3.87 5.43 1.40 3.83 5.15 1.35 3.15 4.61 1.37 p Value 4.6E−5 5.0E−8 0.14 2.5E−4 1.4E−7 0.19 0.0021 1.0E−6 0.16 Lower limit 2.02 2.96 0.890 1.87 2.80 0.862 1.52 2.50 0.882 of 95% CI Upper limit 7.42 9.99 2.22 7.85 9.47 2.10 6.55 8.50 2.14 of 95% CI OR Quartile 4 24.4 36.1 2.60 39.1 34.7 2.51 34.0 32.0 2.50 p Value 0.0017 4.2E−4 3.3E−4 0.010 4.8E−4 5.7E−4 0.014 6.5E−4 7.9E−4 Lower limit 3.32 4.93 1.54 2.38 4.74 1.49 2.07 4.37 1.46 of 95% CI Upper limit 180 265 4.37 642 254 4.24 559 235 4.27 of 95% CI

TABLE 26.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00296 0.0184 0.00282 0.0168 0.00285 0.0162 Average 0.00925 0.152 0.00872 0.145 0.00897 0.141 Stdev 0.0164 0.395 0.0154 0.387 0.0157 0.382 p (t-test) 7.3E−4 0.0019 0.0039 Min 7.09E−7 3.28E−9 7.09E−7 3.28E−9 7.09E−7 3.28E−9 Max 0.102 2.47 0.102 2.47 0.102 2.47 n (Patient) 90 227 79 238 71 246 sCr only Median 0.00328 0.0222 0.00328 0.0220 0.00329 0.0213 Average 0.00870 0.163 0.00881 0.161 0.00872 0.158 Stdev 0.0153 0.408 0.0154 0.406 0.0155 0.402 p (t-test) 1.3E−4 1.6E−4 2.7E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 0.102 2.47 0.102 2.47 0.102 2.47 n (Patient) 106 211 104 213 99 218 UO only Median 0.00764 0.0192 0.00780 0.0175 0.00764 0.0169 Average 0.0920 0.153 0.0981 0.134 0.0990 0.129 Stdev 0.336 0.351 0.350 0.328 0.361 0.315 p (t-test) 0.14 0.37 0.44 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 6.85E−7 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 212 103 194 121 178 137 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.75 0.59 0.73 0.75 0.57 0.72 0.74 0.57 SE 0.029 0.027 0.035 0.030 0.027 0.033 0.031 0.028 0.033 p Value 7.5E−15 0 0.012 3.1E−14 0 0.035 6.9E−12 0 0.026 nCohort 90 106 212 79 104 194 71 99 178 Recovered nCohort 227 211 103 238 213 121 246 218 137 Non-recovered Cutoff 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Quartile 2 Sensitivity 81% 82% 78% 81% 82% 79% 80% 82% 79% Specificity 39% 40% 26% 42% 39% 28% 41% 39% 28% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 60% 64% 59% 59% 63% 56% 57% 62% 55% Specificity 76% 77% 54% 76% 77% 54% 75% 77% 54% Cutoff 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Quartile 4 Sensitivity 34% 36% 36% 33% 36% 33% 32% 35% 32% Specificity 97% 97% 80% 97% 97% 80% 97% 97% 80% OR Quartile 2 2.65 3.09 1.25 2.99 3.00 1.48 2.71 2.89 1.45 p Value 3.8E−4 2.7E−5 0.43 9.5E−5 4.5E−5 0.15 5.6E−4 8.4E−5 0.16 Lower limit 1.55 1.82 0.717 1.73 1.77 0.863 1.54 1.70 0.861 of 95% CI Upper limit 4.53 5.23 2.18 5.19 5.08 2.54 4.77 4.91 2.46 of 95% CI OR Quartile 3 4.71 6.07 1.72 4.51 5.77 1.48 3.95 5.48 1.46 p Value 3.3E−8 3.9E−11 0.026 3.1E−7 1.3E−10 0.091 5.2E−6 7.1E−10 0.098 Lower limit 2.72 3.56 1.07 2.53 3.38 0.939 2.19 3.19 0.932 of 95% CI Upper limit 8.15 10.4 2.77 8.03 9.85 2.34 7.14 9.41 2.28 of 95% CI OR Quartile 4 14.9 19.7 2.27 18.8 19.1 1.96 16.0 17.5 1.93 p Value 7.7E−6 7.6E−7 0.0022 5.8E−5 1.0E−6 0.011 1.5E−4 2.1E−6 0.012 Lower limit 4.56 6.05 1.34 4.49 5.85 1.17 3.83 5.36 1.15 of 95% CI Upper limit 48.6 64.3 3.84 78.4 62.1 3.29 67.0 57.0 3.24 of 95% CI

TABLE 26.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00324 0.0215 0.00306 0.0200 0.00321 0.0178 Average 0.0146 0.165 0.0151 0.160 0.0158 0.153 Stdev 0.0391 0.415 0.0403 0.409 0.0422 0.401 p (t-test) 1.3E−4 3.0E−4 8.6E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 0.361 2.47 0.361 2.47 0.361 2.47 n (Patient) 114 203 107 210 97 220 sCr only Median 0.00365 0.0249 0.00367 0.0232 0.00365 0.0226 Average 0.0139 0.176 0.0142 0.174 0.0142 0.170 Stdev 0.0371 0.427 0.0375 0.424 0.0382 0.420 p (t-test) 2.6E−5 3.9E−5 7.2E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 0.361 2.47 0.361 2.47 0.361 2.47 n (Patient) 127 190 124 193 119 198 UO only Median 0.00730 0.0207 0.00764 0.0182 0.00724 0.0182 Average 0.0934 0.151 0.0971 0.139 0.0964 0.135 Stdev 0.335 0.354 0.342 0.340 0.350 0.328 p (t-test) 0.16 0.30 0.32 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 214 101 204 111 190 125 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.73 0.59 0.72 0.73 0.57 0.71 0.72 0.59 SE 0.028 0.028 0.035 0.029 0.028 0.034 0.029 0.028 0.033 p Value 1.8E−15 0 0.0072 2.2E−14 6.7E−16 0.029 7.5E−13 1.1E−15 0.0092 nCohort 114 127 214 107 124 204 97 119 190 Recovered nCohort 203 190 101 210 193 111 220 198 125 Non-recovered Cutoff 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Quartile 2 Sensitivity 82% 83% 79% 82% 83% 79% 82% 82% 80% Specificity 38% 37% 27% 39% 37% 27% 40% 37% 28% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 63% 65% 61% 61% 64% 58% 60% 64% 58% Specificity 72% 72% 55% 71% 72% 54% 71% 72% 55% Cutoff 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Quartile 4 Sensitivity 36% 38% 34% 35% 38% 32% 33% 37% 32% Specificity 94% 94% 79% 93% 94% 78% 93% 94% 79% OR Quartile 2 2.81 2.90 1.42 3.02 2.86 1.45 3.03 2.73 1.59 p Value 1.1E−4 6.7E−5 0.23 3.8E−5 8.2E−5 0.19 4.4E−5 1.6E−4 0.093 Lower limit 1.67 1.72 0.803 1.79 1.70 0.833 1.78 1.62 0.926 of 95% CI Upper limit 4.74 4.90 2.50 5.11 4.82 2.52 5.15 4.60 2.73 of 95% CI OR Quartile 3 4.28 4.94 1.95 3.83 4.57 1.59 3.63 4.56 1.64 p Value 1.0E−8 1.8E−10 0.0065 1.5E−7 1.2E−9 0.050 9.5E−7 1.9E−9 0.033 Lower limit 2.60 3.02 1.21 2.32 2.80 0.999 2.17 2.78 1.04 of 95% CI Upper limit 7.04 8.07 3.17 6.32 7.46 2.54 6.07 7.48 2.59 of 95% CI OR Quartile 4 8.58 10.7 1.91 7.61 10.2 1.67 6.38 9.34 1.82 p Value 2.5E−7 1.3E−8 0.017 1.1E−6 2.6E−8 0.052 9.1E−6 8.1E−8 0.022 Lower limit 3.79 4.73 1.12 3.36 4.50 0.994 2.82 4.13 1.09 of 95% CI Upper limit 19.4 24.2 3.23 17.2 23.0 2.82 14.5 21.1 3.05 of 95% CI

TABLE 26.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00383 0.0238 0.00365 0.0217 0.00348 0.0194 Average 0.0158 0.181 0.0164 0.173 0.0171 0.164 Stdev 0.0389 0.434 0.0402 0.426 0.0419 0.416 p (t-test) 1.5E−5 5.3E−5 2.0E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 0.361 2.47 0.361 2.47 0.361 2.47 n (Patient) 134 183 125 192 114 203 sCr only Median 0.00383 0.0285 0.00396 0.0284 0.00379 0.0272 Average 0.0150 0.187 0.0152 0.185 0.0151 0.183 Stdev 0.0379 0.440 0.0382 0.438 0.0385 0.435 p (t-test) 5.8E−6 8.0E−6 1.2E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 0.361 2.47 0.361 2.47 0.361 2.47 n (Patient) 140 177 138 179 135 182 UO only Median 0.00717 0.0215 0.00711 0.0199 0.00710 0.0184 Average 0.0925 0.154 0.0956 0.143 0.0948 0.139 Stdev 0.333 0.357 0.340 0.344 0.347 0.333 p (t-test) 0.14 0.24 0.26 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 216 99 207 108 194 121 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.74 0.61 0.71 0.73 0.59 0.70 0.73 0.59 SE 0.028 0.028 0.035 0.029 0.028 0.034 0.029 0.028 0.033 p Value 5.1E−15 0 0.0024 1.1E−13 4.4E−16 0.0073 5.9E−12 0 0.0063 nCohort 134 140 216 125 138 207 114 135 194 Recovered nCohort 183 177 99 192 179 108 203 182 121 Non-recovered Cutoff 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Quartile 2 Sensitivity 83% 84% 80% 83% 84% 80% 82% 84% 79% Specificity 35% 36% 27% 37% 36% 28% 38% 37% 28% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 64% 66% 65% 62% 65% 61% 61% 65% 60% Specificity 69% 70% 56% 68% 70% 56% 68% 70% 56% Cutoff 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Quartile 4 Sensitivity 38% 40% 34% 36% 40% 32% 34% 39% 32% Specificity 93% 94% 79% 92% 93% 79% 91% 93% 79% OR Quartile 2 2.55 3.05 1.48 2.81 2.94 1.49 2.81 3.10 1.48 p Value 4.3E−4 3.8E−5 0.18 1.1E−4 6.2E−5 0.17 1.1E−4 2.7E−5 0.15 Lower limit 1.51 1.79 0.836 1.66 1.73 0.849 1.67 1.83 0.863 of 95% CI Upper limit 4.29 5.18 2.64 4.73 4.98 2.60 4.74 5.27 2.54 of 95% CI OR Quartile 3 4.12 4.55 2.37 3.46 4.31 1.96 3.33 4.44 1.85 p Value 5.7E−9 4.8E−10 5.8E−4 3.0E−7 1.7E−9 0.0053 1.2E−6 8.4E−10 0.0091 Lower limit 2.56 2.82 1.45 2.15 2.68 1.22 2.05 2.78 1.16 of 95% CI Upper limit 6.63 7.33 3.88 5.57 6.94 3.16 5.41 7.24 2.92 of 95% CI OR Quartile 4 7.68 9.75 1.99 6.60 9.42 1.78 5.47 8.95 1.83 p Value 1.8E−8 1.6E−9 0.011 1.9E−7 2.7E−9 0.031 2.7E−6 6.1E−9 0.022 Lower limit 3.78 4.66 1.17 3.24 4.50 1.05 2.69 4.28 1.09 of 95% CI Upper limit 15.6 20.4 3.37 13.4 19.7 3.00 11.1 18.7 3.07 of 95% CI

TABLE 26.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00615 0.0222 0.00601 0.0213 0.00601 0.0213 Average 0.0697 0.172 0.0707 0.168 0.0707 0.168 Stdev 0.294 0.392 0.297 0.388 0.297 0.388 p (t-test) 0.0086 0.012 0.012 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.43 2.47 2.43 2.47 2.43 2.47 n (Patient) 188 129 185 132 185 132 sCr only Median 0.00460 0.0298 0.00458 0.0284 0.00443 0.0284 Average 0.0332 0.191 0.0336 0.187 0.0338 0.186 Stdev 0.161 0.443 0.163 0.439 0.163 0.438 p (t-test) 3.2E−5 4.9E−5 5.7E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 1.92 2.47 1.92 2.47 1.92 2.47 n (Patient) 160 157 157 160 156 161 UO only Median 0.00709 0.0213 0.00710 0.0207 0.00709 0.0209 Average 0.0861 0.164 0.0872 0.160 0.0868 0.159 Stdev 0.328 0.363 0.330 0.359 0.332 0.355 p (t-test) 0.056 0.074 0.075 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 211 104 208 107 205 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.71 0.61 0.65 0.71 0.61 0.65 0.71 0.61 SE 0.032 0.029 0.034 0.032 0.029 0.034 0.032 0.029 0.034 p Value 2.6E−6 3.5E−13 9.2E−4 3.0E−6 3.9E−13 0.0021 3.0E−6 3.8E−13 0.0013 nCohort Recovered 188 160 211 185 157 208 185 156 205 nCohort Non- 129 157 104 132 160 107 132 161 110 recovered Cutoff Quartile 2 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Sensitivity 83% 83% 80% 83% 84% 79% 83% 84% 79% Specificity 30% 33% 27% 31% 34% 27% 31% 34% 27% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 67% 68% 65% 66% 68% 64% 66% 68% 64% Specificity 61% 68% 57% 61% 68% 57% 61% 68% 57% Cutoff Quartile 4 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Sensitivity 36% 41% 35% 35% 40% 34% 35% 40% 34% Specificity 82% 90% 80% 82% 90% 79% 82% 90% 80% OR Quartile 2 2.12 2.50 1.50 2.23 2.63 1.46 2.23 2.67 1.42 Upper limit of 95% CI p Value 0.0080 7.9E−4 0.16 0.0046 4.0E−4 0.19 0.0046 3.2E−4 0.21 Lower limit of 95% 1.22 1.46 0.851 1.28 1.54 0.834 1.28 1.57 0.818 CI 3.68 4.26 2.64 3.87 4.48 2.55 3.87 4.56 2.47 OR Quartile 3 3.15 4.44 2.54 3.03 4.57 2.29 3.03 4.44 2.33 Upper limit of 95% CI p Value 1.6E−6 5.6E−10 1.8E−4 3.0E−6 2.9E−10 7.3E−4 3.0E−6 5.6E−10 5.2E−4 Lower limit of 95% 1.97 2.77 1.56 1.90 2.85 1.41 1.90 2.77 1.44 CI 5.04 7.12 4.13 4.83 7.34 3.69 4.83 7.12 3.75 OR Quartile 4 2.51 6.19 2.07 2.38 5.88 1.95 2.38 5.77 1.97 Upper limit of 95% CI p Value 4.9E−4 3.8E−9 0.0066 0.0010 1.0E−8 0.013 0.0010 1.4E−8 0.011 Lower limit of 95% 1.50 3.38 1.22 1.42 3.20 1.15 1.42 3.15 1.17 CI 4.21 11.4 3.50 3.98 10.8 3.28 3.98 10.6 3.31

Example 27. Use of C—C Motif Chemokine 23 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 23 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 27.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00335 0.0207 0.00326 0.0192 0.00296 0.0192 Average 0.0505 0.143 0.0522 0.140 0.0531 0.138 Stdev 0.299 0.357 0.305 0.354 0.311 0.351 p (t-test) 0.022 0.031 0.039 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 6.85E−7 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 108 209 104 213 100 217 sCr only Median 0.00335 0.0249 0.00335 0.0235 0.00335 0.0231 Average 0.0443 0.156 0.0449 0.154 0.0456 0.153 Stdev 0.277 0.372 0.279 0.370 0.282 0.368 p (t-test) 0.0042 0.0052 0.0063 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 126 190 124 192 122 194 UO only Median 0.00809 0.0175 0.00780 0.0169 0.00744 0.0175 Average 0.0873 0.171 0.0900 0.155 0.0930 0.144 Stdev 0.327 0.370 0.337 0.348 0.345 0.335 p (t-test) 0.048 0.11 0.20 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 6.85E−7 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 222 93 208 107 198 117 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.74 0.57 0.70 0.74 0.58 0.72 0.73 0.59 SE 0.029 0.027 0.036 0.029 0.028 0.034 0.029 0.028 0.034 p Value 9.9E−13 0 0.042 3.1E−12 0 0.021 1.4E−13 0 0.011 nCohort Recovered 108 126 222 104 124 208 100 122 198 nCohort Non- 209 190 93 213 192 107 217 194 117 recovered Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 80% 83% 75% 81% 83% 79% 81% 82% 79% Specificity 35% 37% 25% 37% 37% 27% 38% 37% 28% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 62% 66% 58% 61% 65% 57% 60% 64% 57% Specificity 72% 74% 53% 71% 73% 53% 72% 73% 54% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 36% 38% 39% 35% 38% 38% 35% 38% 38% Specificity 95% 95% 81% 95% 95% 82% 96% 95% 82% OR Quartile 2 2.22 2.94 1.03 2.42 2.84 1.35 2.63 2.75 1.49 Upper limit of 95% CI p Value 0.0027 5.6E−5 0.93 9.9E−4 9.0E−5 0.29 3.3E−4 1.4E−4 0.15 Lower limit of 95% 1.32 1.74 0.586 1.43 1.68 0.773 1.55 1.63 0.863 CI 3.75 4.96 1.80 4.08 4.79 2.34 4.46 4.63 2.57 OR Quartile 3 4.19 5.42 1.57 3.79 5.14 1.52 3.92 4.89 1.58 Upper limit of 95% CI p Value 2.7E−8 2.8E−11 0.070 2.4E−7 1.0E−10 0.082 2.0E−7 3.6E−10 0.053 Lower limit of 95% 2.53 3.30 0.963 2.29 3.13 0.949 2.34 2.98 0.994 CI 6.95 8.91 2.56 6.28 8.45 2.43 6.55 8.02 2.50 OR Quartile 4 11.5 12.5 2.63 10.8 12.1 2.78 12.9 11.7 2.81 Upper limit of 95% CI p Value 3.5E−7 1.3E−8 3.9E−4 7.5E−7 2.1E−8 1.4E−4 1.4E−6 3.2E−8 1.1E−4 Lower limit of 95% 4.50 5.23 1.54 4.20 5.05 1.64 4.58 4.88 1.66 CI 29.5 29.8 4.48 27.6 28.8 4.70 36.5 27.9 4.73

TABLE 27.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00396 0.0222 0.00396 0.0220 0.00368 0.0217 Average 0.0487 0.158 0.0494 0.157 0.0503 0.153 Stdev 0.268 0.380 0.270 0.378 0.275 0.374 p (t-test) 0.0045 0.0055 0.0081 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 6.85E−7 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 136 181 134 183 129 188 sCr only Median 0.00420 0.0272 0.00413 0.0260 0.00413 0.0259 Average 0.0452 0.170 0.0455 0.169 0.0460 0.167 Stdev 0.257 0.392 0.257 0.391 0.259 0.389 p (t-test) 0.0011 0.0012 0.0015 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 149 168 148 169 146 171 UO only Median 0.00730 0.0220 0.00744 0.0215 0.00722 0.0209 Average 0.0877 0.173 0.0889 0.165 0.0927 0.149 Stdev 0.326 0.374 0.330 0.363 0.340 0.344 p (t-test) 0.045 0.069 0.17 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 6.85E−7 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 226 89 220 95 207 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.69 0.72 0.61 0.69 0.72 0.61 0.69 0.71 0.61 SE 0.029 0.028 0.036 0.029 0.028 0.035 0.029 0.029 0.034 p Value 1.5E−10 3.1E−14 0.0017 2.5E−10 3.3E−14 0.0019 3.4E−11 1.9E−13 0.0020 nCohort Recovered 136 149 226 134 148 220 129 146 207 nCohort Non- 181 168 89 183 169 95 188 171 108 recovered Cutoff Quartile 2 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Sensitivity 81% 83% 80% 81% 83% 81% 81% 82% 81% Specificity 32% 34% 27% 33% 34% 28% 34% 34% 29% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 63% 67% 64% 62% 66% 62% 62% 65% 60% Specificity 67% 68% 55% 66% 68% 55% 67% 68% 55% Cutoff Quartile 4 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Sensitivity 38% 40% 38% 37% 40% 38% 37% 39% 36% Specificity 91% 91% 80% 91% 91% 80% 91% 91% 81% OR Quartile 2 2.00 2.42 1.46 2.07 2.46 1.64 2.26 2.37 1.75 Upper limit of 95% CI p Value 0.0086 9.7E−4 0.21 0.0058 7.7E−4 0.10 0.0020 0.0012 0.054 Lower limit of 95% 1.19 1.43 0.804 1.23 1.46 0.908 1.35 1.41 0.990 CI 3.34 4.09 2.64 3.46 4.16 2.97 3.80 4.00 3.11 OR Quartile 3 3.44 4.34 2.20 3.27 4.22 2.00 3.22 4.00 1.85 Upper limit of 95% CI p Value 2.2E−7 1.0E−9 0.0022 6.6E−7 2.0E−9 0.0057 1.1E−6 7.0E−9 0.011 Lower limit of 95% 2.16 2.71 1.33 2.05 2.64 1.22 2.01 2.50 1.15 CI 5.49 6.95 3.66 5.21 6.76 3.28 5.15 6.39 2.97 OR Quartile 4 6.22 6.94 2.49 6.01 6.82 2.51 6.22 6.59 2.36 Upper limit of 95% CI p Value 7.0E−8 4.5E−9 9.1E−4 1.2E−7 6.1E−9 6.9E−4 1.7E−7 1.1E−8 0.0013 Lower limit of 95% 3.20 3.63 1.45 3.09 3.57 1.48 3.13 3.45 1.40 CI 12.1 13.3 4.26 11.7 13.0 4.27 12.3 12.6 3.98

TABLE 27.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00427 0.0260 0.00427 0.0259 0.00396 0.0232 Average 0.0490 0.170 0.0496 0.168 0.0506 0.164 Stdev 0.255 0.397 0.257 0.395 0.260 0.391 p (t-test) 0.0015 0.0019 0.0029 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 154 163 152 165 148 169 sCr only Median 0.00427 0.0295 0.00427 0.0293 0.00427 0.0289 Average 0.0471 0.177 0.0473 0.176 0.0476 0.175 Stdev 0.250 0.404 0.251 0.403 0.251 0.402 p (t-test) 6.0E−4 6.8E−4 7.9E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 161 156 160 157 159 158 UO only Median 0.00710 0.0222 0.00711 0.0221 0.00717 0.0212 Average 0.0895 0.167 0.0899 0.165 0.0934 0.150 Stdev 0.328 0.370 0.328 0.368 0.336 0.351 p (t-test) 0.068 0.075 0.17 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 224 91 223 92 212 103 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.72 0.62 0.69 0.70 0.62 0.70 0.72 0.61 SE 0.029 0.029 0.036 0.029 0.029 0.036 0.029 0.029 0.035 p Value 7.9E−13 7.8E−15 4.9E−4 3.3E−12 8.7E−15 6.1E−4 2.7E−12 5.9E−14 0.0013 nCohort Recovered 154 161 224 152 160 223 148 159 212 nCohort Non- 163 156 91 165 157 92 169 158 103 recovered Cutoff Quartile 2 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Sensitivity 84% 85% 81% 84% 853% 82% 84% 85% 83% Specificity 34% 35% 28% 34% 35% 28% 35% 35% 29% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 65% 67% 67% 64% 67% 66% 63% 66% 62% Specificity 66% 66% 57% 65% 66% 57% 65% 66% 56% Cutoff Quartile 4 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Sensitivity 39% 41% 37% 39% 41% 37% 38% 41% 35% Specificity 90% 90% 80% 89% 90% 80% 89% 90% 80% OR Quartile 2 2.77 3.08 1.67 2.66 3.14 1.70 2.85 2.95 1.91 p Value 1.9E−4 5.7E−5 0.097 3.1E−4 4.4E−5 0.085 1.2E−4 9.5E−5 0.032 Lower limit of 95% 1.62 1.78 0.912 1.56 1.81 0.930 1.67 1.71 1.06 CI Upper limit of 95% 4.72 5.34 3.04 4.52 5.43 3.10 4.85 5.09 3.44 CI OR Quartile 3 3.54 4.08 2.66 3.36 3.96 2.56 3.19 3.85 2.06 p Value 8.4E−8 3.9E−9 1.7E−4 2.7E−7 7.3E−9 2.9E−4 8.0E−7 14E−8 000033 Lower limit of 95% 2.23 2.56 1.60 2.12 2.49 1.54 2.01 2.41 1.27 CI Upper limit of 95% 5.63 6.51 4.44 5.32 6.32 4.24 5.05 6.14 3.33 CI OR Quartile 4 5.58 6.30 2.3 5.39 6.19 2.32 5.03 6.09 2.11 p Value 2.7E−8 2.7E−9 0.0016 5.0E−8 3.8E−9 0.0021 1.7E−7 5.4E−9 0.0053 Upper limit of 95% 3.04 3.44 1.39 2.94 3.38 1.36 2.75 3.32 1.25 CI Lower limit of 95% 10.2 11.6 4.06 9.87 11.4 3.96 9.21 11.2 3.57 CI

TABLE 27.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine smples for the “recovered” and “non-recoevered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine iutput RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00443 0.0285 0.00428 0.0272 0.00427 0.0249 Average 0.0480 0.183 0.0488 0.179 0.0497 0.175 Stdev 0.245 0.412 0.248 0.409 0.251 0.405 p (t-test) 4.1E−4 6.1E−4 0.0010 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 168 149 165 152 161 156 sCr only Median 0.00443 0.0336 0.00428 0.0317 0.00443 0.0298 Average 0.0468 0.188 0.0471 0.186 0.0473 0.185 Stdev 0.243 0.417 0.243 0.416 0.244 0.415 p (t-test) 2.2E−4 2.5E−4 2.9E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 172 145 171 146 170 147 UO only Median 0.00711 0.0227 0.00710 0.0222 0.00710 0.0215 Average 0.0898 0.165 0.0902 0.164 0.0927 0.153 Stdev 0.328 0.368 0.329 0.366 0.335 0.354 p (t-test) 0.074 0.081 0.15 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 223 92 222 93 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.71 0.72 0.62 0.70 0.72 0.62 0.70 0.72 0.62 SE 0.029 0.029 0.036 0.029 0.029 0.036 0.029 0.029 0.035 p Value 8.1E−13 LOE−14 4.6E−4 3.6E−12 L2E−14 4.9E−4 3.5E−12 8.0E−14 8.7E−4 nCohort Recovered 168 172 223 165 171 222 161 170 214 nCohort Non- 149 145 92 152 146 93 156 147 101 recovered Cutoff Quartile 2 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Sensitivity 85% 86% 82% 84% 86% 82% 85% 86% 82% Specificity 33% 34% 28% 33% 35% 28% 34% 34% 29% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 67% 68% 68% 66% 68% 68% 65% 67% 64% Specificity 65% 65% 57% 64% 65% 57% 64% 65% 57% Cutoff Quartile 4 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Sensitivity 41% 43% 37% 40% 42% 37% 39% 42% 35% Specificity 89% 90% 80% 88% 89% 80% 88% 89% 79% OR Quartile 2 2.74 3.26 1.70 2.67 3.32 1.73 2.85 3.11 1.84 Upper limit of 95% CI p Value 3.1E−4 4.4E−5 0.085 4.0E−4 3.4E−5 0.074 1.6E−4 7.3E−5 0.043 Lower limit of 95% 1.58 1.85 0.930 1.55 1.88 0.949 1.66 1.77 1.02 CI 4.74 5.76 3.10 4.59 5.85 3.16 4.91 5.44 3.32 OR Quartile 3 3.77 4.02 2.93 3.46 3.90 2.81 3.26 3.78 2.35 Upper limit of 95% CI p Value 2.4E−8 6.5E−9 4.2E−5 1.5E−7 1.2E−8 7.2E−5 4.7E−7 2.3E−8 6.1E−4 Lower limit of 95% 2.37 2.51 1.75 2.18 2.44 1.69 2.06 2.37 1.44 CI 6.01 6.43 4.89 5.49 6.22 4.67 5.16 6.03 3.83 OR Quartile 4 5.44 6.39 2.32 5.15 6.27 2.27 4.80 6.16 2.05 Upper limit of 95% CI p Value 9.7E−9 6.6E−10 0.0021 2.7E−8 9.5E−10 0.0027 9.8E−8 1.4E−9 0.0077 Lower limit of 95% 3.05 3.55 1.36 2.89 3.48 1.33 2.70 3.42 1.21 CI 9.69 11.5 3.96 9.18 11.3 3.87 8.54 11.1 3.47

TABLE 27.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine smples for the “recovered” and “non-recoevered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine iutput RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00638 0.0249 0.00638 0.0235 0.00651 0.0232 Average 0.0834 0.161 0.0842 0.158 0.0846 0.157 Stdev 0.333 0.350 0.334 0.348 0.335 0.347 p (t-test) 0.052 0.063 0.069 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 203 114 201 116 200 117 sCr only Median 0.00574 0.0344 0.00574 0.0344 0.00574 0.0344 Average 0.0509 0.196 0.0509 0.196 0.0509 0.196 Stdev 0.238 0.433 0.238 0.433 0.238 0.433 p (t-test) 1.7E−4 1.7E−4 1.7E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 185 132 185 132 185 132 UO only Median 0.00708 0.0235 0.00701 0.0232 0.00709 0.0222 Average 0.0838 0.174 0.0841 0.172 0.0849 0.169 Stdev 0.324 0.372 0.325 0.370 0.326 0.367 p (t-test) 0.029 0.033 0.041 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 217 98 216 99 214 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.71 0.64 0.64 0.71 0.64 0.64 0.71 0.63 SE 0.033 0.030 0.035 0.033 0.030 0.035 0.033 0.030 0.035 p Value 4.7E−6 7.9E−12 6.6E−5 LIE−5 7.9E−12 7.0E−5 1.5E−5 7.9E−12 1.9E−4 nCohort Recovered 203 185 217 201 185 216 200 185 214 nCohort Non- 114 132 98 116 132 99 117 132 101 recovered Cutoff Quartile 2 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 0.00202 0.00202 0.00207 Sensitivity 83% 84% 83% 83% 84% 83% 83% 84% 82% Specificity 30% 31% 29% 29% 31% 29% 30% 31% 29% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 69% 71% 69% 68% 71% 69% 68% 71% 67% Specificity 61% 65% 59% 60% 65% 58% 60% 65% 58% Cutoff Quartile 4 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 0.0477 0.0477 0.0478 Sensitivity 37% 42% 37% 36% 42% 36% 36% 42% 36% Specificity 81% 87% 80% 81% 87% 80% 81% 87% 80% OR Quartile 2 2.10 2.41 1.91 1.99 2.41 1.94 2.03 2.41 1.84 p Value 0.012 0.0021 0.035 0.017 0.0021 0.030 0.015 0.0021 0.043 Lower limit of 95% 1.18 1.38 1.05 1.13 1.38 1.07 1.15 1.38 1.02 CI Upper limit of 95% 3.74 4.23 3.47 3.52 4.23 3.54 3.59 4.23 3.32 CI OR Quartile 3 3.47 4.57 3.20 3.23 4.57 3.07 3.12 4.57 2.84 p Value 5.7E−7 7.0E−10 7.1E−6 1.9E−6 7.0E−10 1.3E−5 3.3E−6 7.0E−10 3.8E−5 Lower limit of 95% 2.13 2.82 1.93 1.99 2.82 1.86 1.93 2.82 1.73 CI Upper limit of 95% 5.65 7.40 5.31 5.23 7.40 5.08 5.04 7.40 4.66 CI OR Quartile 4 2.53 4.94 2.35 2.43 4.94 2.30 2.39 4.94 2.20 p Value 4.5E−4 1.3E−8 0.0016 7.6E−4 1.3E−8 0.0020 9.7E−4 1.3E−8 0.0033 Lower limit of 95% 1.51 2.85 1.38 1.45 2.85 1.36 1.42 2.85 1.30 CI Upper limit of 95% 4.26 8.57 3.99 4.09 8.57 3.90 4.00 8.57 3.73 CI

Example 28. Use of C—C Motif Chemokine 23 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 23 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 28.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine smples for the “recovered” and “non-recoevered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine iutput RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00597 0.0390 0.00601 0.0499 0.00709 0.0479 Average 0.0619 0.201 0.0592 0.231 0.0780 0.208 Stdev 0.283 0.412 0.273 0.438 0.320 0.380 p (t-test) 4.5E−4 3.0E−5 0.0029 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 1.82 n (Patient) 205 112 221 96 236 81 sCr only Median 0.00599 0.0390 0.00630 0.0479 0.00694 0.0443 Average 0.0644 0.202 0.0621 0.230 0.0812 0.199 Stdev 0.284 0.417 0.274 0.444 0.323 0.379 p (t-test) 6.2E−4 5.8E−5 0.0072 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 1.82 n (Patient) 208 108 223 93 235 81 UO only Median 0.00870 0.0349 0.00864 0.0832 0.00859 0.0861 Average 0.0897 0.265 0.0887 0.286 0.0884 0.294 Stdev 0.311 0.484 0.309 0.497 0.309 0.502 p (t-test) 0.0023 9.0E−4 6.2E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 275 40 278 37 279 36 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.70 0.70 0.63 0.73 0.72 0.66 0.70 0.69 0.68 SE 0.032 0.032 0.050 0.032 0.033 0.051 0.036 0.036 0.051 p Value 9.9E−11 L6E−10 0.011 6.3E−13 2.0E−11 0.0017 4.1E−8 LIE−7 4.7E−4 nCohort Non- 205 208 275 221 223 278 236 235 279 persistent nCohort Persistent 112 108 40 96 93 37 81 81 36 Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 84% 85% 78% 86% 87% 81% 84% 86% 83% Specificity 30% 30% 25% 30% 30% 26% 28% 29% 26% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 72% 73% 68% 76% 75% 73% 73% 74% 75% Specificity 62% 62% 52% 61% 61% 53% 58% 58% 53% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 47% 47% 48% 52% 51% 51% 51% 48% 53% Specificity 87% 87% 78% 86% 86% 78% 83% 83% 78% OR Quartile 2 2.21 2.50 1.18 2.72 2.90 1.50 2.03 2.59 1.77 p Value 0.0080 0.0031 0.69 0.0026 0.0019 0.36 0.035 0.0073 0.22 Lower limit of 95% 1.23 1.36 0.534 1.42 1.48 0.630 1.05 1.29 0.709 CI Upper limit of 95% 3.98 4.59 2.59 5.22 5.67 3.56 3.92 5.19 4.43 CI OR Quartile 3 4.25 4.45 2.28 4.98 4.67 3.03 3.65 3.99 3.39 p Value 1.5E−8 9.3E−9 0.021 6.0E−9 2.5E−8 0.0044 4.6E−6 1.3E−6 0.0025 Lower limit of 95% 2.58 2.67 1.13 2.90 2.71 1.41 2.10 2.28 1.54 CI Upper limit of 95% 7.02 7.40 4.61 8.56 8.03 6.50 6.34 7.00 7.47 CI OR Quartile 4 5.92 5.75 3.24 6.92 6.10 3.84 5.18 4.53 4.08 p Value 2.2E−10 4.2E−10 7.4E−4 8.7E−12 1.4E−10 1.9E−4 6.3E−9 8.7E−8 1.1E−4 Lower limit of 95% 3.42 3.32 1.64 3.97 3.51 1.89 2.97 2.60 2.00 CI Upper limit of 95% 10.3 9.96 6.42 12.1 10.6 7.76 9.02 7.87 8.33 CI

TABLE 28.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine smples for the “recovered” and “non-recoevered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine iutput RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00500 0.0347 0.00576 0.0465 0.00630 0.0472 Average 0.0512 0.196 0.0487 0.224 0.0681 0.207 Stdev 0.245 0.429 0.234 0.456 0.294 0.413 p (t-test) 1.6E−4 8.6E−6 7.0E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.34 n (Patient) 186 131 204 113 219 98 sCr only Median 0.00576 0.0338 0.00578 0.0422 0.00615 0.0387 Average 0.0548 0.199 0.0526 0.222 0.0724 0.198 Stdev 0.244 0.439 0.236 0.461 0.297 0.412 p (t-test) 2.1E−4 2.2E−5 0.0023 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 1.82 n (Patient) 192 124 206 110 218 98 UO only Median 0.00780 0.0298 0.00780 0.0355 0.00778 0.0410 Average 0.0894 0.210 0.0882 0.224 0.0879 0.228 Stdev 0.319 0.414 0.317 0.425 0.316 0.428 p (t-test) 0.0014 0.0073 0.0059 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 256 59 260 55 261 54 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.72 0.71 0.65 0.74 0.73 0.67 0.72 0.70 0.68 SE 0.030 0.031 0.042 0.030 0.031 0.043 0.033 0.033 0.043 p Value 2.1E−13 1.3E−11 4.3E−4 6.7E−16 8.0E−14 8.4E−5 2.1E−11 8.4E−10 2.3E−5 nCohort Non- 186 192 256 204 206 260 219 218 261 persistent nCohort Persistent 131 124 59 113 110 55 98 98 54 Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 84% 85% 81% 87% 87% 84% 86% 87% 85% Specificity 31% 31% 27% 31% 32% 27% 30% 30% 27% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 72% 72% 71% 76% 75% 75% 74% 74% 76% Specificity 65% 64% 55% 64% 64% 55% 61% 61% 55% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 45% 44% 46% 50% 47% 47% 50% 46% 48% Specificity 89% 87% 80% 88% 87% 80% 86% 84% 80% OR Quartile 2 2.37 2.51 1.58 2.99 3.16 1.88 2.53 2.84 2.15 p Value 0.0025 0.0017 0.21 5.3E−4 3.7E−4 0.10 0.0042 0.0017 0.061 Lower limit of 95% 1.36 1.41 0.775 1.61 1.68 0.876 1.34 1.48 0.967 CI Upper limit of 95% 4.16 4.47 3.22 5.54 5.95 4.05 4.78 5.45 4.78 CI OR Quartile 3 4.73 4.53 29.98 5.72 5.37 3.58 4.52 4.57 3.88 p Value 3.5E−10 1.5E−9 5.0E−4 4.4E−11 2.2E−10 13E−4 2.3E−8 1.9E−8 7.2E−5 Lower limit of 95% 2.91 2.78 1.61 3.40 3.20 1.86 2.66 2.69 1.99 CI Upper limit of 95% 7.68 7.40 5.51 9.60 9.02 6.88 7.66 7.76 7.58 CI OR Quartile 4 6.44 5.15 3.31 7.37 5.94 3.50 6.06 4.59 3.64 p Value 1.5E−10 5.2E−9 8.2E−5 3.7E−12 2.3E−10 5.5E−5 1.2E−10 3.1E−8 3.6E−5 Lower limit of 95% 3.64 2.97 1.82 4.19 3.43 1.90 3.50 2.68 1.97 CI Upper limit of 95% 11.4 8.93 6.01 12.9 10.3 6.44 10.5 7.89 6.73 CI

TABLE 28.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) pnly, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00463 0.0344 0.00466 0.0443 0.00597 0.0434 Average 0.0451 0.193 0.0427 0.214 0.0589 0.207 Stdev 0.240 0.421 0.230 0.440 0.280 0.415 p (t-test) 1.1E−4 9.0E−6 1.9E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.34 n (Patient) 175 142 190 127 205 112 Cr only Median 0.00500 0.0344 0.00531 0.0425 0.00599 0.0387 Average 0.0479 0.198 0.0462 0.217 0.0622 0.206 Stdev 0.239 0.430 0.231 0.449 0.280 0.421 p (t-test) 9.9E−5 1.2E−5 3.3E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.34 n (Patient) 1182 134 195 121 208 108 UO only Median 0.00765 0.0238 0.00751 0.0272 0.00751 0.0272 Average 0.0863 0.203 0.0849 0.218 0.0849 0.218 Stdev 0.314 0.415 0.311 0.428 0.311 0.428 p (t-test) 0.012 0.0053 0.0053 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 246 69 251 64 251 64 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.73 0.72 0.63 0.75 0.75 0.66 0.72 0.71 0.66 SE 0.029 0.029 0.040 0.029 0.029 0.040 0.031 0.032 0.040 p Value 7.8E−15 2.9E−14 9.5E−4 0 0 9.0E−5 9.3E−13 1.4E−11 9.0E−5 nCohort 175 182 246 190 195 251 205 208 251 Non- persistent nCohort 142 134 69 127 121 64 112 108 64 Persistent Cutoff 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Quartile 2 Sensitivity 84% 85% 81% 87% 88% 84% 86% 87% 84% Specificity 32% 32% 27% 33% 33% 27% 31% 31% 27% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 71% 72% 68% 76% 75% 72% 74% 74% 72% Specificity 67% 66% 55% 67% 66% 55% 63% 62% 55% Cutoff 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Quartile 4 Sensitivity 44% 44% 42% 48% 47% 44% 47% 45% 44% Specificity 90% 89% 80% 90% 89% 80% 87% 86% 80% OR 2.43 2.73 1.58 3.13 3.45 2.05 2.66 3.05 2.05 Quartile 2 p Value 0.0015 5.1E−4 0.18 1.6E−4 8.5E−5 0.054 0.0016 5.6E−4 0.054 Lower 1.41 1.55 0.811 1.73 1.86 0.987 1.45 1.62 0.987 limit of 95% CI Upper 4.21 4.82 3.07 5.68 6.40 4.25 4.88 5.75 4.25 limit of 95% CI OR 4.97 4.89 2.60 6.24 5.80 3.17 4.86 4.76 3.17 Quartile 3 p Value 6.1E−11 1.4E−10 9.3E−4 1.2E−12 1.2E−11 1.6E−4 1.2E−9 2.6E−9 1.6E−4 Lower 3.07 3.01 1.48 3.77 3.49 1.74 2.92 2.85 1.74 limit of 95% CI Upper 8.03 7.94 4.57 10.3 9.63 5.77 8.08 7.96 5.77 limit of 95% CI OR 7.41 6.37 2.84 8.32 7.00 3.05 5.92 4.93 3.05 Quartile 4 p Value 6.0E−11 3.0E−10 3.3E−4 1.7E−12 2.1E−11 1.7E−4 2.2E−10 7.8E−9 1.7E−4 Lower 4.07 3.58 1.61 4.62 3.96 1.70 3.42 2.87 1.70 limit of 95% CI Upper 13.5 11.3 5.03 15.0 12.4 5.46 10.3 8.47 5.46 limit of 95% CI

TABLE 28.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00463 0.0338 0.00470 0.0434 0.00578 0.0422 Average 0.0456 0.190 0.0429 0.212 0.0478 0.221 Stdev 0.241 0.418 0.231 0.439 0.227 0.458 p (t-test) 1.5E−4 1.1E−5 9.8E−6 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 173 144 189 128 201 116 sCr only Median 0.00552 0.0338 0.00552 0.0422 0.00599 0.0387 Average 0.0484 0.195 0.0462 0.212 0.0508 0.222 Stdev 0.240 0.428 0.233 0.444 0.228 0.465 p (t-test) 1.4E−4 1.9E−5 1.6E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 180 136 192 124 204 112 UO only Median 0.00751 0.0235 0.00751 0.0272 0.00751 0.0272 Average 0.0865 0.192 0.0847 0.207 0.0847 0.207 Stdev 0.318 0.400 0.314 0.413 0.314 0.413 p (t-test) 0.019 0.0080 0.0080 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 239 76 245 70 245 70 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.72 0.72 0.63 0.75 0.74 0.66 0.72 0.71 0.66 SE 0.029 0.030 0.038 0.029 0.029 0.039 0.031 0.031 0.039 p Value 1.7E−14 3.4E−13 4.9E−4 0 2.2E−16 3.8E−5 2.7E−13 7.6E−12 3.8E−5 nCohort 173 180 239 189 192 245 201 204 245 Non- persistent nCohort 144 136 76 128 124 70 116 112 70 Persistent Cutoff 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Quartile 2 Sensitivity 84% 85% 83% 87% 87% 86% 86% 87% 86% Specificity 32% 32% 28% 33% 33% 28% 31% 31% 28% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 70% 71% 67% 75% 74% 71% 74% 73% 71% Specificity 66% 66% 55% 67% 66% 56% 64% 63% 56% Cutoff 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Quartile 4 Sensitivity 44% 43% 41% 48% 47% 43% 47% 46% 43% Specificity 90% 89% 80% 90% 89% 80% 87% 86% 80% OR 2.52 2.60 1.85 3.19 3.30 2.35 2.85 2.96 2.35 Quartile 2 p Value 9.5E−4 8.2E−4 0.068 1.3E−4 1.1E−4 0.021 7.0E−4 6.0E−4 0.021 Lower 1.46 1.49 0.955 1.76 1.80 1.14 1.56 1.59 1.14 limit of 95% CI Upper 4.36 4.56 3.58 5.77 6.04 4.86 5.23 5.49 4.86 limit of 95% CI OR 4.66 4.57 2.52 6.00 5.49 3.17 5.03 4.60 3.17 Quartile 3 p Value 2.5E−10 5.6E−10 8.5E−4 2.5E−12 2.6E−11 8.8E−5 3.8E−10 3.2E−9 8.8E−5 Lower 2.89 2.83 1.46 3.63 3.33 1.78 3.03 2.78 1.78 limit of 95% CI Upper 7.50 7.38 4.33 9.91 9.05 5.65 8.33 7.63 5.65 limit of 95% CI OR 7.14 6.13 2.74 8.15 7.16 3.00 5.86 5.26 3.00 Quartile 4 p Value 1.3E−10 6.6E−10 3.8E−4 2.6E−12 1.9E−11 1.5E−4 3.0E−10 2.5E−9 1.5E−4 Lower 3.92 3.45 1.57 4.53 4.03 1.70 3.38 3.05 1.70 limit of 95% CI Upper 13.0 10.9 4.78 14.7 12.7 5.29 10.2 9.07 5.29 limit of 95% CI

TABLE 28.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00463 0.0317 0.00466 0.0384 0.00578 0.0369 Average 0.0464 0.185 0.0438 0.205 0.0485 0.214 Stdev 0.244 0.413 0.234 0.432 0.229 0.452 p (t-test) 2.6E−4 2.7E−5 2.1E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 169 148 184 133 197 120 sCr only Median 0.00552 0.0317 0.00552 0.0369 0.00599 0.0351 Average 0.0493 0.190 0.0469 0.206 0.0516 0.215 Stdev 0.243 0.423 0.235 0.438 0.231 0.458 p (t-test) 2.5E−4 3.8E−5 3.5E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 176 140 188 128 200 116 UO only Median 0.00717 0.0285 0.00722 0.0291 0.00722 0.0291 Average 0.0798 0.205 0.0791 0.212 0.0791 0.212 Stdev 0.312 0.402 0.311 0.409 0.311 0.409 p (t-test) 0.0045 0.0028 0.0028 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 234 81 237 78 237 78 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.72 0.71 0.66 0.74 0.73 0.66 0.72 0.70 0.66 SE 0.029 0.030 0.037 0.029 0.030 0.037 0.031 0.031 0.037 p Value 7.2E−14 2.8E−12 2.1E−5 0 4.7E−15 1.1E−5 2.3E−12 7.2E−11 1.1E−5 nCohort 169 176 234 184 188 237 197 200 237 Non- persistent nCohort 148 140 81 133 128 78 120 116 78 Persistent Cutoff 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Quartile 2 Sensitivity 84% 84% 84% 86% 86% 85% 86% 85% 85% Specificity 33% 32% 28% 33% 32% 28% 31% 31% 28% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 69% 69% 69% 73% 73% 71% 72% 72% 71% Specificity 66% 65% 56% 66% 65% 57% 63% 62% 57% Cutoff 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Quartile 4 Sensitivity 43% 42% 42% 46% 45% 42% 45% 44% 42% Specificity 90% 89% 81% 90% 89% 81% 87% 86% 81% OR 2.49 2.37 2.05 3.17 2.94 2.17 2.78 2.62 2.17 Quartile 2 p Value 9.7E−4 0.0020 0.032 1.1E−4 3.0E−4 0.025 7.4E−4 0.0015 0.025 Lower 1.45 1.37 1.06 1.77 1.64 1.10 1.54 1.44 1.10 limit of 95% CI Upper 4.29 4.11 3.97 5.68 5.27 4.27 5.04 4.75 4.27 limit of 95% CI OR 4.36 4.25 2.90 5.30 5.03 3.11 4.58 4.19 3.11 Quartile 3 p Value 9.9E−10 2.3E−9 1.0E−4 2.4E−11 1.2E−10 5.3E−5 1.7E−9 1.4E−8 5.3E−5 Lower 2.72 2.65 1.69 3.25 3.08 1.79 2.79 2.56 1.79 limit of 95% CI Upper 6.99 6.84 4.96 8.65 8.22 5.39 7.51 6.87 5.39 limit of 95% CI OR 6.63 5.68 3.04 7.36 6.59 3.04 5.38 4.82 3.04 Quartile 4 p Value 5.6E−10 3.0E−9 7.1E−5 2.2E−11 1.0E−10 7.8E−5 1.7E−9 1.3E−8 7.8E−5 Lower 3.64 3.20 1.76 4.10 3.72 1.75 3.11 2.80 1.75 limit of 95% CI Upper 12.0 10.1 5.26 13.2 11.7 5.29 9.30 8.29 5.29 limit of 95% CI

Example 29. Use of C—C Motif Chemokine 23 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 23 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 29.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00330 0.0226 0.00434 0.0230 0.00458 0.0289 Average 0.0467 0.150 0.0475 0.175 0.0486 0.192 Stdev 0.285 0.365 0.250 0.402 0.239 0.426 p (t-test) 0.0087  8.3E−4  1.7E−4 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 119 198 158 159 179 138 sCr only Median 0.00367 0.0249 0.00428 0.0259 0.00460 0.0351 Average 0.0479 0.156 0.0473 0.179 0.0487 0.199 Stdev 0.277 0.374 0.250 0.406 0.238 0.433 p (t-test) 0.0055  5.6E−4  9.8E−5 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 130 186 162 154 184 132 UO only Median 0.00834 0.0202 0.00834 0.0367 0.00834 0.0554 Average 0.0863 0.195 0.0833 0.236 0.0824 0.261 Stdev 0.317 0.404 0.308 0.442 0.304 0.464 p (t-test) 0.016 0.0018  5.1E−4 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 241 74 256 59 263 52 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.73 0.60 0.69 0.71 0.63 0.71 0.72 0.66 SE 0.028 0.028 0.039 0.030 0.029 0.042 0.030 0.030 0.044 p Value  4.4E−16 2.2E−16 0.0089  2.1E−10  1.2E−12 0.0019  2.9E−12  2.0E−13 2.9E−4 nCohort Non- 119 130 241 158 162 256 179 184 263 persistent nCohort Persistent 198 186 74 159 154 59 138 132 52 Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 82% 83% 78% 82% 83% 76% 85% 86% 79% Specificity 36% 36% 26% 32% 33% 25% 32% 33% 26% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 64% 66% 61% 65% 67% 66% 67% 69% 69% Specificity 72% 72% 53% 65% 66% 54% 63% 64% 54% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 37% 38% 41% 39% 40% 47% 42% 43% 52% Specificity 95% 94% 80% 89% 90% 80% 88% 88% 80% OR Quartile 2 2.55 2.73 1.28 2.08 2.39 1.09 2.67 2.88 1.30 p Value 4.3E−4 1.7E−4  0.43 0.0063 0.0014 0.79 5.9E−4 3.2E−4 0.48 Lower limit of 1.51 1.62 0.688 1.23 1.40 0.564 1.53 1.62 0.632 95% CI Upper limit of 4.28 4.60 2.39 3.50 4.08 2.12 4.68 5.12 2.67 95% CI OR Quartile 3 4.56 4.98 1.76 3.35 3.93 2.24 3.54 3.88 2.60 p Value 1.9E−9 1.2E−10 0.037 2.7E−7 9.5E−9 0.0075 1.2E−7 2.3E−8 0.0033 Lower limit of 2.78 3.05 1.03 2.11 2.46 1.24 2.22 2.41 1.38 95% CI Upper limit of 7.48 8.12 2.99 5.31 6.27 4.06 5.65 6.24 4.92 95% CI OR Quartile 4 11.2 9.42 2.67 4.97 5.75 3.63 5.17 5.60 4.38 p Value 5.0E−8 1.4E−8  5.9E−4 7.8E−8 9.3E−9 2.2E−5 8.7E−9 2.0E−9 3.3E−6 Lower limit of 4.71 4.34 1.53 2.77 3.16 2.00 2.96 3.19 2.35 95% CI Upper limit of 26.8 20.4 4.68 8.93 10.4 6.59 9.06 9.83 8.17 95% CI

TABLE 29.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00314 0.0220 0.00406 0.0249 0.00406 0.0317 Average 0.0484 0.146 0.0463 0.167 0.0462 0.182 Stdev 0.294 0.360 0.258 0.390 0.247 0.409 p (t-test) 0.015 0.0015  3.6E−4 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 112 205 147 170 165 152 sCr only Median 0.00335 0.0249 0.00413 0.0272 0.00420 0.0341 Average 0.0485 0.153 0.0481 0.172 0.0484 0.184 Stdev 0.281 0.371 0.256 0.397 0.247 0.413 p (t-test) 0.0074 0.0012  3.9E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 126 190 154 162 169 147 UO only Median 0.00730 0.0215 0.00744 0.0285 0.00737 0.0326 Average 0.0880 0.169 0.0843 0.197 0.0833 0.212 Stdev 0.328 0.367 0.318 0.396 0.314 0.412 p (t-test) 0.055 0.011 0.0053 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 222 93 238 77 245 70 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.74 0.61 0.71 0.72 0.64 0.73 0.74 0.66 SE 0.028 0.027 0.036 0.029 0.028 0.038 0.028 0.028 0.039 p Value  2.2E−16 0 0.0013  2.4E−13  8.0E−15 1.9E−4  2.2E−16 0 2.7E−5 nCohort Non- 112 126 222 147 154 238 165 169 245 persistent nCohort 205 190 93 170 162 77 152 147 70 Persistent Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 82% 83% 81% 82% 84% 79% 86% 86% 81% Specificity 38% 37% 27% 33% 34% 26% 35% 35% 27% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 63% 65% 63% 66% 67% 69% 68% 69% 71% Specificity 73% 73% 55% 68% 68% 56% 67% 67% 56% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 37% 38% 39% 39% 40% 44% 42% 42% 47% Specificity 96% 94% 81% 90% 90% 81% 90% 90% 81% OR Quartile 2 2.72 2.94 1.58 2.33 2.74 1.37 3.12 3.41 1.62 p Value 1.7E−4 5.6E−5 0.13 0.0015 2.2E−4 0.32 5.8E−5 2.3E−5  0.16 Lower limit of 1.62 1.74 0.873 1.38 1.61 0.737 1.79 1.93 0.831 95% CI Upper limit of 4.59 4.96 2.86 3.93 4.69 2.55 5.43 6.01 3.14 95% CI OR Quartile 3 4.64 5.08 2.16 4.11 4.41 2.80 4.33 4.57 3.17 p Value 2.6E−9  1.1E−10 0.0025 3.7E−9  7.3E−10 2.2E−4 1.0E−9 3.5E−10 8.8E−5 Lower limit of 2.80 3.10 1.31 2.57 2.75 1.62 2.71 2.84 1.78 95% CI Upper limit of 7.69 8.33 3.55 6.57 7.06 4.83 6.94 7.35 5.65 95% CI OR Quartile 4 12.3 10.4 2.63 6.03 6.05 3.39 6.77 6.52 3.86 p Value 1.6E−7 2.0E−8 3.9E−4 2.4E−8 1.2E−8 1.6E−5  6.9E−10 8.3E−10 3.2E−6 Lower limit of 4.82 4.58 1.54 3.21 3.26 1.95 3.69 3.58 2.19 95% CI Upper limit of 31.6 23.5 4.48 11.3 11.2 5.91 12.4 11.9 6.81 95% CI

TABLE 29.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00321 0.0217 0.00424 0.0232 0.00424 0.0285 Average 0.0488 0.145 0.0472 0.164 0.0478 0.174 Stdev 0.295 0.359 0.261 0.388 0.252 0.401 p (t-test) 0.016 0.0022  8.8E−4 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 111 206 144 173 158 159 sCr only Median 0.00352 0.0235 0.00427 0.0259 0.00427 0.0293 Average 0.0493 0.152 0.0490 0.169 0.0500 0.177 Stdev 0.283 0.369 0.259 0.394 0.252 0.407 p (t-test) 0.0091 0.0018  8.9E−4 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 124 192 151 165 163 153 UO only Median 0.00717 0.0212 0.00722 0.0235 0.00717 0.0260 Average 0.0910 0.155 0.0880 0.172 0.0869 0.182 Stdev 0.336 0.351 0.327 0.371 0.322 0.384 p (t-test) 0.12 0.050 0.029 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 212 103 225 90 232 83 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.73 0.61 0.70 0.71 0.62 0.72 0.72 0.64 SE 0.028 0.028 0.035 0.029 0.029 0.036 0.028 0.029 0.037 p Value  6.7E−16 2.2E−16 0.0019  3.1E−12  4.3E−13 5.8E−4  8.4E−15  3.6E−14 1.3E−4 nCohort Non- 111 124 212 144 151 225 158 163 232 persistent nCohort Persistent 206 192 103 173 165 90 159 153 83 Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 82% 83% 81% 82% 83% 80% 85% 85% 82% Specificity 38% 37% 28% 33% 34% 27% 35% 34% 28% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 63% 65% 63% 65% 66% 67% 67% 67% 69% Specificity 73% 73% 56% 67% 68% 56% 66% 66% 56% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 36% 38% 37% 38% 39% 40% 40% 41% 42% Specificity 95% 94% 81% 90% 90% 81% 90% 90% 81% OR Quartile 2 2.78 2.84 1.60 2.29 2.50 1.49 3.00 2.96 1.73 p Value 1.3E−4 9.0E−5  0.11 0.0018 6.9E−4 0.19 7.4E−5 1.1E−4 0.089 Lower limit of 1.65 1.68 0.902 1.36 1.47 0.821 1.74 1.71 0.921 95% CI Upper limit of 4.69 4.79 2.84 3.85 4.23 2.70 5.17 5.12 3.24 95% CI OR Quartile 3 4.52 4.83 2.19 3.79 4.05 2.59 3.96 4.05 2.84 p Value 4.7E−9 3.9E−10 0.0015 2.3E−8 5.0E−9 2.6E−4 7.4E−9 5.0E−9 1.2E−4 Lower limit of 2.73 2.95 1.35 2.37 2.54 1.55 2.48 2.53 1.67 95% CI Upper limit of 7.50 7.90 3.55 6.05 6.48 4.32 6.32 6.46 4.84 95% CI OR Quartile 4 12.1 10.0 2.44 5.73 5.75 2.82 5.98 5.85 3.12 p Value 2.0E−7 3.2E−8  8.9E−4 5.9E−8 3.0E−8 1.5E−4 7.4E−9 6.6E−9 4.4E−5 Lower limit of 4.74 4.43 1.44 3.05 3.10 1.65 3.26 3.22 1.81 95% CI Upper limit of 31.1 22.7 4.12 10.8 10.7 4.83 11.0 10.6 5.38 95% CI

TABLE 29.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00314 0.0215 0.00420 0.0231 0.00424 0.0284 Average 0.0492 0.144 0.0475 0.164 0.0484 0.172 Stdev 0.296 0.358 0.262 0.387 0.254 0.399 p (t-test) 0.018 0.0024 0.0011 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 110 207 143 174 156 161 sCr only Median 0.00352 0.0235 0.00424 0.0249 0.00427 0.0289 Average 0.0493 0.152 0.0493 0.168 0.0503 0.176 Stdev 0.283 0.369 0.259 0.393 0.253 0.405 p (t-test) 0.0091 0.0020 0.0010 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 124 192 150 166 162 154 UO only Median 0.00710 0.0207 0.00717 0.0222 0.00717 0.0232 Average 0.0925 0.150 0.0897 0.163 0.0888 0.170 Stdev 0.339 0.345 0.330 0.363 0.326 0.373 p (t-test) 0.16 0.079 0.056 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 208 107 220 95 226 89 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.73 0.73 0.61 0.70 0.71 0.62 0.71 0.72 0.63 SE 0.028 0.028 0.034 0.029 0.029 0.035 0.029 0.029 0.036 p Value 6.7E−16 2.2E−16 0.0020 4.7E−12 6.7E−13 6.9E−4 7.8E−14 6.0E−14 3.6E−4 nCohort Non- 110 124 208 143 150 220 156 162 226 persistent nCohort Persistent 207 192 107 174 166 95 161 154 89 Cutoff Quartile 2 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Sensitivity 82% 83% 81% 82% 83% 81% 84% 85% 82% Specificity 38% 37% 28% 345 34% 28% 35% 35% 28% Cutoff Quartile 3 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Sensitivity 62% 65% 63% 64% 66% 66% 66% 67% 67% Specificity 73% 73% 56% 67% 67% 57% 66% 66% 57% Cutoff Quartile 4 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Sensitivity 36% 38% 36% 38% 39% 38% 40% 40% 39% Specificity 95% 94% 80% 90% 90% 80% 90% 90% 81% OR Quartile 2 2.84 2.84 1.72 2.33 2.54 1.64 2.88 3.01 1.76 p Value 9.5E−5 9.0E−5 0.062 0.0015 5.5E−4 0.10 1.2E−4 8.3E−5 0.070 Lower limit of 1.68 1.68 0.972 1.38 1.50 0.908 1.68 1.74 0.954 95% CI Upper limit of 4.79 4.79 3.05 3.92 4.31 2.97 4.94 5.21 3.26 95% CI OR Quartile 3 4.41 4.83 2.15 3.69 3.94 2.59 3.75 3.93 2.70 p Value 8.5E−9 3.9E−10 0.0017 4.2E−8 9.3E−9 2.0E−4 2.5E−8 9.5E−9 1.6E−4 Lower limit of 2.66 2.95 1.34 2.31 2.47 1.57 2.35 2.46 1.61 95% CI Upper limit of 7.31 7.90 3.47 5.89 6.29 4.28 5.96 6.27 4.52 95% CI OR Quartile 4 11.9 10.0 2.24 5.63 5.65 2.51 5.77 5.75 2.68 p Value 2.4E−7 3.2E−8 0.0025 7.9E−8 4.1E−8 6.9E−4 1.4E−8 9.3E−9 3.3E−4 Lower limit of 4.66 4.43 1.33 3.00 3.04 1.48 3.15 3.16 1.57 95% CI Upper limit of 30.6 22.7 3.78 10.6 10.5 4.27 10.6 10.4 4.59 95% CI

TABLE 29.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UOQ) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 0.00294 0.0212 0.00396 0.0222 0.00413 0.0259 Average 0.0503 0.142 0.0487 0.159 0.0498 0.166 Stdev 0.302 0.355 0.267 0.382 0.259 0.393 p (t-test) 0.024 0.0039 0.0022 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 106 211 138 179 150 167 sCr only Median 0.00330 0.0232 0.00396 0.0249 0.00413 0.0289 Average 0.0499 0.150 0.0498 0.164 0.0508 0.172 Stdev 0.287 0.366 0.263 0.389 0.256 0.401 p (t-test) 0.011 0.0028 0.0015 Min 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 3.28E−9 Max 2.47 2.43 2.47 2.43 2.47 2.43 n (Patient) 121 195 146 170 158 158 UO only Median 0.00675 0.0215 0.00709 0.0232 0.00708 0.0249 Average 0.0858 0.158 0.0841 0.169 0.0836 0.175 Stdev 0.335 0.350 0.327 0.364 0.324 0.372 p (t-test) 0.070 0.038 0.028 Min 6.85E−7 3.28E−9 6.85E−7 3.28E−9 3.28E−9 3.28E−9 Max 2.47 1.82 2.47 1.82 2.47 1.82 n (Patient) 202 113 212 103 217 98 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.73 0.74 0.63 0.70 0.72 0.63 0.71 0.72 0.64 SE 0.028 0.028 0.033 0.029 0.028 0.034 0.029 0.028 0.035 p Value 0 0 1.5E−4 2.8E−12 3.9E−14 1.1E−4 2.3E−13 3.6E−15 3.8E−5 nCohort 106 121 202 138 146 212 150 158 217 Non- persistent nCohort 211 195 113 179 170 103 167 158 98 Persistent Cutoff 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 0.00202 0.00200 0.00207 Quartile 2 Sensitivity 82% 83% 82% 83% 84% 82% 84% 85% 83% Specificity 40% 38% 29% 35% 35% 28% 35% 35% 29% Cutoff 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 0.0104 Quartile 3 Sensitivity 63% 65% 65% 64% 66% 67% 65% 68% 68% Specificity 75% 74% 58% 68% 69% 58% 67% 68% 58% Cutoff 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 0.0477 0.0468 0.0478 Quartile 4 Sensitivity 36% 37% 35% 37% 38% 37% 38% 39% 38% Specificity 95% 94% 81% 90% 90% 81% 89% 89% 81% OR 3.09 3.01 1.92 2.55 2.72 1.75 2.96 3.22 1.91 Quartile 2 p Value 2.7E−5 3.8E−5 0.025 4.5E−4 2.1E−4 0.060 7.1E−5 3.0E−5 0.035 Lower limit 1.82 1.78 1.08 1.51 1.60 0.976 1.73 1.86 1.05 of 95% CI Upper limit 5.23 5.09 3.39 4.29 4.62 3.12 5.06 5.58 3.47 of 95% CI OR 4.89 5.42 2.51 3.84 4.45 2.80 3.76 4.40 2.99 Quartile 3 p Value 2.0E−9 4.6E−11 1.5E−4 2.1E−8 6.8E−10 4.1E−5 2.5E−8 7.3E−10 2.0E−5 Lower limit 2.91 3.28 1.56 2.40 2.77 1.71 2.36 2.75 1.81 of 95% CI Upper limit 8.21 8.97 4.04 6.15 7.15 4.59 5.99 7.05 4.95 of 95% CI OR 11.1 9.53 2.29 5.17 5.27 2.44 5.20 5.36 2.53 Quartile 4 p Value 5.2E−7 6.3E−8 0.0018 3.2E−7 1.3E−7 8.9E−4 9.1E−8 3.4E−8 6.0E−4 Lower limit 4.35 4.21 1.36 2.75 2.84 1.44 2.84 2.95 1.49 of 95% CI Upper limit 28.6 21.6 3.85 9.72 9.78 4.12 9.53 9.72 4.29 of 95% CI

Example 30. Use of C—C Motif Chemokine 24 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 24 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is risk of injury (R), injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 30.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UOQ) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 9.06 33.1 10.0 32.0 11.0 31.8 Average 45.2 80.7 42.4 79.7 44.4 78.8 Stdev 111 153 104 152 110 151 p (t-test) 0.098 0.11 0.16 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 670 1150 670 1150 670 1150 n (Patient) 57 259 46 270 41 275 sCr only Median 11.9 35.0 11.5 36.2 10.0 37.4 Average 54.4 79.7 54.2 79.5 52.8 79.5 Stdev 147 146 149 145 152 144 p (t-test) 0.19 0.19 0.18 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 76 239 74 241 70 245 UO only Median 19.3 47.2 18.3 43.7 18.4 40.8 Average 52.4 109 49.3 104 44.3 103 Stdev 115 182 113 175 79.7 186 p (t-test) 8.4E−4 8.6E−4 3.2E−4 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 670 1150 n (Patient) 193 121 172 142 155 159 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.65 0.62 0.66 0.64 0.63 0.66 0.63 0.65 0.64 SE 0.037 0.035 0.032 0.041 0.035 0.031 0.043 0.035 0.031 p Value 8.8E−5 4.2E−4 3.1E−7 8.5E−4 1.5E−4 1.3E−7 0.0027 1.7E−5 8.6E−6 nCohort 57 76 193 46 74 172 41 70 155 Recovered nCohort 259 239 121 270 241 142 275 245 159 Non- recovered Cutoff 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Quartile 2 Sensitivity 78% 79% 87% 77% 79% 85% 77% 79% 82% Specificity 40% 37% 33% 39% 38% 34% 39% 40% 33% Cutoff 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Quartile 3 Sensitivity 54% 55% 65% 53% 56% 63% 53% 56% 60% Specificity 68% 66% 60% 67% 68% 61% 68% 70% 61% Cutoff 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Quartile 4 Sensitivity 28% 28% 39% 27% 28% 36% 27% 28% 35% Specificity 88% 84% 83% 87% 85% 84% 85% 86% 85% OR Quartile 2.45 2.15 3.18 2.20 2.27 2.93 2.15 2.54 2.29 2 p Value 0.0037 0.0073 1.8E−4 0.019 0.0043 1.7E−4 0.029 0.0013 0.0021 Lower limit 1.34 1.23 1.73 1.14 1.29 1.67 1.08 1.44 1.35 of 95% CI Upper limit 4.50 3.76 5.83 4.25 3.98 5.14 4.28 4.48 3.89 of 95% CI OR Quartile 2.55 2.37 2.77 2.33 2.61 2.71 2.40 2.96 2.35 3 p Value 0.0026 0.0017 2.3E−5 0.012 6.2E−4 2.0E−5 0.014 1.9E−4 2.2E−4 Lower limit 1.39 1.39 1.73 1.20 1.51 1.71 1.19 1.67 1.49 of 95% CI Upper limit 4.69 4.06 4.45 4.51 4.52 4.29 4.83 5.23 3.69 of 95% CI OR Quartile 2.75 2.08 3.20 2.47 2.25 2.88 2.11 2.35 2.89 4 p Value 0.018 0.035 1.5E−5 0.049 0.023 9.2E−5 0.11 0.021 1.3E−4 Lower limit 1.19 1.05 1.89 1.01 1.12 1.70 0.852 1.14 1.68 of 95% CI Upper limit 6.35 4.09 5.41 6.07 4.53 4.90 5.22 4.86 4.97 of 95% CI

TABLE 30.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the ″recovered″ and ″non recovered″ cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UOQ) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.5 38.0 12.2 33.1 12.2 33.1 Average 34.3 90.3 37.0 86.7 37.2 85.1 Stdev 77.1 164 81.7 161 85.6 158 p (t-test) 0.0020 0.0088 0.015 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 670 1150 670 1150 670 1150 n (Patient) 90 226 79 237 71 245 sCr only Median 13.6 38.0 12.9 38.4 11.7 39.2 Average 47.3 88.0 47.0 87.7 45.5 87.5 Stdev 125 154 127 154 129 152 p (t-test) 0.019 0.020 0.018 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 106 210 104 212 99 217 UO only Median 22.7 51.1 20.4 49.9 21.0 43.2 Average 52.2 119 50.8 111 46.6 110 Stdev 112 193 113 183 83.6 196 p (t-test) 1.2E−4 3.3E−4 1.4E−4 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 670 1150 n (Patient) 211 103 193 121 177 137 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.65 0.64 0.67 0.63 0.64 0.66 0.63 0.66 0.64 SE 0.032 0.032 0.034 0.034 0.032 0.032 0.035 0.031 0.032 p Value 1.9E−6 2.1E−5 3.8E−7 1.2E−4 7.4E−6 4.5E−7 1.6E−4 4.3E−7 1.8E−5 nCohort 90 106 211 79 104 193 71 99 177 Recovered nCohort 226 210 103 237 212 121 245 217 137 Non- recovered Cutoff 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Quartile 2 Sensitivity 80% 80% 86% 78% 81% 85% 78% 81% 82% Specificity 37% 36% 31% 35% 37% 32% 37% 38% 31% Cutoff 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Quartile 3 Sensitivity 56% 57% 66% 54% 57% 64% 54% 58% 61% Specificity 66% 63% 58% 63% 64% 59% 65% 67% 58% Cutoff 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Quartile 4 Sensitivity 31% 31% 43% 30% 31% 39% 29% 31% 37% Specificity 91% 87% 83% 90% 88% 83% 90% 89% 84% OR Quartile 2.27 2.30 2.83 2.00 2.40 2.64 2.09 2.67 2.12 2 p Value 0.0029 0.0018 0.0013 0.014 0.0011 0.0011 0.011 2.7E−4 0.0066 Lower limit 1.32 1.36 1.50 1.15 1.42 1.47 1.18 1.58 1.23 of 95% CI Upper limit 3.88 3.89 5.34 3.49 4.06 4.75 3.70 4.54 3.66 of 95% CI OR Quartile 2.44 2.25 2.66 2.06 2.41 2.47 2.18 2.72 2.14 3 p Value 5.7E−4 9.5E−4 9.2E−5 0.0069 3.8E−4 1.5E−4 0.0052 8.1E−5 0.0010 Lower limit 1.47 1.39 1.63 1.22 1.48 1.55 1.26 1.65 1.36 of 95% CI Upper limit 4.06 3.63 4.35 3.48 3.91 3.95 3.78 4.47 3.37 of 95% CI OR Quartile 4.70 2.95 3.75 3.80 3.16 3.20 3.81 3.65 3.16 4 p Value 9.9E−5 8.4E−4 1.2E−6 8.3E−4 5.1E−4 1.5E−5 0.0015 2.3E−4 2.3E−5 Lower limit 2.16 1.56 2.20 1.74 1.65 1.89 1.66 1.83 1.85 of 95% CI Upper limit 10.2 5.55 6.39 8.30 6.06 5.41 8.70 7.27 5.37 of 95% CI

TABLE 30.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UOQ) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Recovered Non-recovered Recovered Non-recovered Recovered Non-recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 17.2 39.4 15.8 37.8 12.9 38.2 Average 42.9 92.0 44.2 89.7 42.7 88.3 Stdev 85.0 169 87.6 167 89.9 164 p (t-test) 0.0040 0.0086 0.010 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 670 1150 670 1150 670 1150 n (Patient) 114 202 107 209 97 219 sCr only Median 17.6 40.2 16.4 40.3 14.3 41.4 Average 49.6 90.9 49.7 90.2 47.1 90.8 Stdev 119 160 120 159 122 157 p (t-test) 0.014 0.016 0.0100 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 127 189 124 192 119 197 UO only Median 20.4 54.9 20.3 51.1 20.3 49.9 Average 53.0 119 53.5 112 54.5 104 Stdev 114 192 117 185 120 176 p (t-test) 1.8E−4 6.9E−4 0.0033 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 213 101 203 111 189 125 Recovery Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.63 0.63 0.68 0.63 0.63 0.67 0.64 0.65 0.65 SE 0.032 0.031 0.033 0.032 0.031 0.033 0.032 0.031 0.032 p Value 4.4E−5 5.7E−5 4.6E−8 9.2E−5 3.0E−5 2.3E−7 3.2E−5 7.6E−7 1.9E−6 nCohort 114 127 213 107 124 203 97 119 189 Recovered nCohort 202 189 101 209 192 111 219 197 125 Non- recovered Cutoff 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Quartile 2 Sensitivity 80% 81% 86% 80% 81% 86% 80% 82% 85% Specificity 34% 34% 31% 36% 35% 31% 36% 36% 32% Cutoff 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Quartile 3 Sensitivity 57% 58% 68% 56% 58% 67% 56% 59% 64% Specificity 62% 61% 59% 62% 62% 59% 64% 65% 59% Cutoff 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Quartile 4 Sensitivity 32% 32% 43% 31% 32% 40% 31% 32% 38% Specificity 87% 85% 83% 86% 85% 83% 88% 87% 84% OR Quartile 2.11 2.18 2.73 2.26 2.30 2.67 2.25 2.53 2.59 2 p Value 0.0049 0.0032 0.0020 0.0023 0.0016 0.0015 0.0028 4.7E−4 0.0012 Lower limit of 95% CI 1.25 1.30 1.45 1.34 1.37 1.46 1.32 1.50 1.46 Upper limit 3.54 3.65 5.15 3.81 3.86 4.90 3.81 4.26 4.62 of 95% CI OR Quartile 2.18 2.17 3.06 2.05 2.25 2.89 2.27 2.63 2.59 3 p Value 0.0011 9.5E−4 1.2E−5 0.0032 6.1E−4 1.7E−5 0.0011 5.9E−5 6.6E−5 Lower limit of 95% CI 1.36 1.37 1.86 1.27 1.41 1.78 1.39 1.64 1.62 Upper limit 3.49 3.43 5.05 3.29 3.56 4.69 3.72 4.21 4.12 of 95% CI OR Quartile 3.06 2.64 3.65 2.71 2.74 3.15 3.12 3.34 3.18 4 p Value 3.9E−4 9.4E−4 2.0E−6 0.0016 7.2E−4 1.9E−5 8.6E−4 1.3E−4 1.7E−5 Lower limit of 95% CI 1.65 1.49 2.14 1.46 1.53 1.86 1.60 1.80 1.87 Upper limit 5.68 4.70 6.21 5.03 4.92 5.34 6.10 6.19 5.38 of 95% CI

TABLE 30.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 17.9 40.3 16.9 38.2 15.9 39.4 Average 53.8 89.4 55.9 86.4 45.0 90.8 Stdev 124 159 128 156 91.0 168 p (t-test) 0.032 0.070 0.0073 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 670 1150 n (Patient) 134 182 125 191 114 202 sCr only Median 17.2 40.6 16.4 40.6 15.8 41.4 Average 51.7 92.3 51.8 91.8 49.7 92.7 Stdev 119 163 120 162 120 161 p (t-test) 0.014 0.016 0.0097 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 140 176 138 178 135 181 UO only Median 20.3 55.1 20.4 52.8 20.3 50.6 Average 57.4 111 58.4 104 54.5 106 Stdev 133 167 136 162 119 178 p (t-test) 0.0027 0.0082 0.0026 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 215 99 206 108 193 121 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.63 0.63 0.69 0.62 0.64 0.67 0.64 0.65 0.66 SE 0.031 0.031 0.033 0.031 0.031 0.033 0.031 0.031 0.032 p Value 2.9E−5 1.6E−5 1.0E−8 9.7E-5 1.1E−5 3.1E−7 7.4E−6 8.2E−7 6.9E−7 nCohort Recovered 134 140 215 125 138 206 114 135 193 nCohort Non- 182 176 99 191 178 108 202 181 121 recovered Cutoff Quartile 2 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Sensitivity 82% 82% 88% 82% 83% 86% 81% 83% 86% Specificity 34% 34% 31% 35% 35% 31% 36% 36% 32% Cutoff Quartile 3 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Sensitivity 58% 59% 70% 57% 59% 67% 57% 60% 64% Specificity 61% 61% 59% 61% 62% 59% 63% 63% 59% Cutoff Quartile 4 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Sensitivity 32% 32% 43% 31% 33% 41% 32% 34% 39% Specificity 85% 84% 83% 84% 85% 83% 87% 87% 83% OR Quartile 2 2.36 2.44 3.28 2.42 2.53 2.79 2.42 2.67 2.90 p Value 0.0012 8.0E−4 5.0E−4 8.4E−4 5.0E−4 0.0012 8.5E−4 2.3E−4 4.6E−4 Lower limit 1.40 1.45 1.68 1.44 1.50 1.50 1.44 1.58 1.60 of 95% CI Upper limit 3.97 4.11 6.41 4.07 4.26 5.19 4.08 4.50 5.25 of 95% CI OR Quartile 3 2.20 2.30 3.32 2.06 2.31 2.85 2.31 2.52 2.62 p Value 7.0E−4 3.2E−4 3.6E−6 0.0020 3.2E−4 2.5E−5 5.0E−4 8.1E−5 6.0E−5 Lower limit 1.39 1.46 2.00 1.30 1.46 1.75 1.44 1.59 1.64 of 95% CI Upper limit 3.47 3.62 5.51 3.26 3.64 4.63 3.71 3.98 4.19 of 95% CI OR Quartile 4 2.73 2.57 3.82 2.35 2.69 3.36 3.06 3.30 3.20 p Value 5.1E−4 8.4E−4 9.2E−7 0.0033 5.3E−4 7.1E−6 3.9E−4 6.1E−5 1.5E−5 Lower limit 1.55 1.48 2.24 1.33 1.54 1.98 1.65 1.84 1.89 of 95% CI Upper limit 4.82 4.47 6.52 4.14 4.72 5.70 5.68 5.93 5.41 of 95% CI

TABLE 30.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 19.2 44.7 19.3 44.7 19.3 44.7 Average 58.6 97.0 59.3 95.2 59.3 95.2 Stdev 138 155 139 154 139 154 p (t-test) 0.022 0.031 0.031 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 187 129 184 132 184 132 sCr only Median 18.4 42.2 19.7 41.6 19.2 41.8 Average 52.9 96.0 53.8 94.3 53.0 94.8 Stdev 117 168 118 167 118 167 p (t-test) 0.0087 0.014 0.011 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 159 157 156 160 155 161 UO only Median 20.4 54.2 20.4 53.9 20.4 52.5 Average 58.5 106 59.0 104 59.0 102 Stdev 135 164 136 163 137 161 p (t-test) 0.0068 0.011 0.012 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 210 104 207 107 204 110 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.64 0.68 0.65 0.63 0.67 0.65 0.63 0.67 SE 0.032 0.031 0.033 0.032 0.031 0.033 0.032 0.031 0.033 p Value 4.0E−7 1.0E−5 7.1E−8 2.2E−6 6.0E−5 2.6E−7 2.2E−6 2.7E−5 1.6E−7 nCohort Recovered 187 159 210 184 156 207 184 155 204 nCohort Non- 129 157 104 132 160 107 132 161 110 recovered Cutoff Quartile 2 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Sensitivity 86% 83% 88% 85% 82% 87% 85% 82% 87% Specificity 33% 33% 31% 32% 32% 31% 32% 32% 32% Cutoff Quartile 3 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Sensitivity 65% 62% 68% 64% 61% 67% 64% 61% 67% Specificity 60% 62% 59% 60% 61% 59% 60% 61% 59% Cutoff Quartile 4 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Sensitivity 37% 35% 41% 36% 34% 40% 36% 35% 40% Specificity 83% 85% 83% 83% 85% 83% 83% 85% 83% OR Quartile 2 2.99 2.34 3.21 2.64 2.13 3.04 2.64 2.17 3.21 p Value 2.4E−4 0.0017 4.4E−4 7.9E−4 0.0047 5.9E−4 7.9E−4 0.0038 3.1E−4 Lower limit 1.66 1.38 1.67 1.50 1.26 1.61 1.50 1.28 1.70 of 95% CI Upper limit 5.36 3.98 6.15 4.66 3.60 5.73 4.66 3.66 6.04 of 95% CI OR Quartile 3 2.85 2.60 3.10 2.75 2.40 2.95 2.75 2.46 3.00 p Value 1.0E−5 3.7E−5 7.7E−6 1.8E−5 1.5E−4 1.5E−5 1.8E−5 9.4E−5 9.8E−6 Lower limit 1.79 1.65 1.89 1.73 1.53 1.81 1.73 1.57 1.84 of 95% CI Upper limit 4.54 4.09 5.10 4.37 3.77 4.82 4.37 3.87 4.87 of 95% CI OR Quartile 4 2.98 3.03 3.41 2.82 2.88 3.19 2.82 3.06 3.22 p Value 4.6E−5 6.4E−5 5.9E−6 1.1E−4 1.4E−4 1.6E−5 1.1E−4 6.5E−5 1.4E−5 Lower limit 1.76 1.76 2.01 1.67 1.67 1.88 1.67 1.77 1.90 of 95% CI Upper limit 5.04 5.23 5.79 4.76 4.96 5.41 4.76 5.30 5.45 of 95% CI

TABLE 30.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 330 375 245 375 364 374 Average 391 540 385 536 399 532 Stdev 281 543 298 536 310 533 p (t-test) 0.19 0.21 0.29 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1070 4690 1070 4690 1070 4690 n (Patient) 25 126 21 130 19 132 sCr only Median 330 374 287 375 304 374 Average 428 535 423 535 433 531 Stdev 402 536 408 534 422 530 p (t-test) 0.32 0.30 0.38 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 29 121 28 122 26 124 UO only Median 364 380 330 384 337 380 Average 494 528 499 519 456 563 Stdev 552 427 573 407 341 632 p (t-test) 0.69 0.81 0.19 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1880 4690 1880 1540 4690 n (Patient) 91 59 84 66 77 73 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.58 0.58 0.54 0.59 0.59 0.55 0.58 0.59 0.54 SE 0.060 0.057 0.049 0.064 0.057 0.048 0.067 0.059 0.047 p Value 0.18 0.14 0.44 0.14 0.097 0.32 0.23 0.13 0.40 nCohort Recovered 25 29 91 21 28 84 19 26 77 nCohort Non- 126 121 59 130 122 66 132 124 73 recovered Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 76% 77% 76% 76% 77% 79% 77% 77% 77% Specificity 32% 34% 26% 33% 36% 29% 37% 38% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 52% 51% 53% 52% 52% 53% 51% 51% 52% Specificity 56% 55% 52% 57% 57% 52% 53% 54% 52% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 27% 26% 29% 26% 26% 26% 26% 26% 26% Specificity 84% 79% 77% 81% 79% 75% 79% 77% 75% OR Quartile 2 1.51 1.75 1.15 1.60 1.87 1.49 1.90 2.14 1.24 p Value 0.39 0.21 0.72 0.36 0.17 0.31 0.22 0.095 0.58 Lower limit 0.591 0.729 0.539 0.592 0.773 0.697 0.689 0.875 0.590 of 95% CI Upper limit 3.84 4.19 2.46 4.31 4.50 3.17 5.25 5.25 2.59 of 95% CI OR Quartile 3 1.36 1.29 1.18 1.42 1.42 1.24 1.15 1.20 1.17 p Value 0.49 0.54 0.62 0.46 0.40 0.51 0.78 0.67 0.62 Lower limit 0.572 0.573 0.614 0.559 0.622 0.651 0.437 0.516 0.618 of 95% CI Upper limit 3.22 2.92 2.28 3.59 3.26 2.37 3.00 2.81 2.23 of 95% CI OR Quartile 4 1.94 1.38 1.35 1.51 1.30 1.04 1.30 1.16 1.07 p Value 0.25 0.52 0.43 0.49 0.60 0.92 0.66 0.77 0.85 Lower limit 0.621 0.515 0.640 0.473 0.485 0.496 0.404 0.428 0.514 of 95% CI Upper limit 6.06 3.69 2.84 4.79 3.50 2.18 4.19 3.14 2.24 of 95% CI

TABLE 30.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 347 376 344 376 344 374 Average 454 537 458 533 471 527 Stdev 394 548 411 540 433 532 p (t-test) 0.38 0.44 0.59 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 40 111 36 115 32 119 sCr only Median 327 380 324 386 291 380 Average 434 548 431 549 436 544 Stdev 392 552 396 549 410 543 p (t-test) 0.21 0.20 0.26 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 44 107 43 108 40 111 UO only Median 350 394 332 396 337 392 Average 480 563 481 551 444 591 Stdev 530 452 548 426 329 663 p (t-test) 0.35 0.42 0.078 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1880 4690 1880 1540 4690 n (Patient) 100 50 93 57 85 65 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.55 0.58 0.56 0.55 0.59 0.57 0.55 0.59 0.56 SE 0.052 0.050 0.050 0.054 0.050 0.049 0.056 0.051 0.048 p Value 0.35 0.097 0.23 0.32 0.073 0.13 0.37 0.073 0.24 nCohort Recovered 40 44 100 36 43 93 32 40 85 nCohort Non- 111 107 50 115 108 57 119 111 65 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity 76% 78% 78% 76% 78% 81% 76% 78% 77% Specificity 28% 32% 27% 28% 33% 29% 31% 35% 27% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity 51% 52% 54% 51% 53% 56% 51% 52% 54% Specificity 52% 55% 52% 53% 56% 54% 53% 55% 53% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity 27% 28% 34% 26% 28% 30% 25% 27% 29% Specificity 80% 82% 79% 78% 81% 77% 75% 80% 78% OR Quartile 2 1.18 1.61 1.31 1.20 1.69 1.71 1.48 1.95 1.24 p Value 0.69 0.23 0.51 0.68 0.19 0.19 0.37 0.098 0.58 Lower limit 0.521 0.740 0.588 0.514 0.772 0.772 0.626 0.885 0.584 of 95% CI Upper limit 2.67 3.52 2.92 2.78 3.70 3.79 3.49 4.31 2.62 of 95% CI OR Quartile 3 1.17 1.32 1.27 1.18 1.41 1.49 1.19 1.34 1.31 p Value 0.68 0.44 0.49 0.67 0.34 0.24 0.66 0.43 0.41 Lower limit 0.566 0.652 0.644 0.556 0.694 0.767 0.545 0.647 0.687 of 95% CI Upper limit 2.41 2.66 2.51 2.49 2.87 2.89 2.61 2.76 2.51 of 95% CI OR Quartile 4 1.48 1.75 1.94 1.24 1.68 1.46 1.01 1.48 1.43 p Value 0.38 0.21 0.087 0.64 0.24 0.32 0.98 0.38 0.34 Lower limit 0.614 0.731 0.908 0.508 0.701 0.690 0.411 0.614 0.685 of 95% CI Upper limit 3.57 4.20 4.13 3.01 4.04 3.08 2.49 3.57 3.00 of 95% CI

TABLE 30.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 369 374 327 380 327 376 Average 469 538 457 541 463 534 Stdev 383 565 394 555 414 543 p (t-test) 0.44 0.36 0.45 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 50 101 46 105 40 111 sCr only Median 327 380 311 392 298 394 Average 447 553 436 557 430 556 Stdev 386 568 389 563 395 556 p (t-test) 0.22 0.17 0.16 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 54 97 52 99 49 102 UO only Median 331 397 329 397 334 390 Average 474 572 477 559 493 527 Stdev 534 443 544 432 563 419 p (t-test) 0.26 0.34 0.68 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1880 4690 1880 4690 1880 n (Patient) 98 52 94 56 86 64 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.53 0.57 0.58 0.55 0.59 0.58 0.55 0.60 0.54 SE 0.050 0.048 0.050 0.050 0.048 0.049 0.052 0.048 0.048 p Value 0.60 0.14 0.10 0.31 0.067 0.12 0.30 0.039 0.37 nCohort Recovered 50 54 98 46 52 94 40 49 86 nCohort Non- 101 97 52 105 99 56 111 102 64 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity 74% 77% 79% 75% 78% 79% 77% 78% 77% Specificity 24% 30% 28% 26% 31% 28% 30% 33% 27% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity 50% 53% 58% 52% 54% 59% 52% 54% 55% Specificity 50% 54% 54% 54% 56% 55% 55% 57% 53% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity 27% 28% 33% 27% 28% 30% 26% 28% 28% Specificity 78% 80% 79% 78% 81% 78% 78% 82% 77% OR Quartile 2 0.911 1.44 1.42 1.07 1.56 1.40 1.40 1.76 1.19 p Value 0.82 0.35 0.39 0.86 0.25 0.40 0.41 0.14 0.65 Lower limit of 0.414 0.676 0.637 0.485 0.731 0.641 0.626 0.824 0.563 95% CI Upper limit of 2.00 3.05 3.15 2.37 3.31 3.06 3.14 3.77 2.52 95% CI OR Quartile 3 1.02 1.29 1.61 1.31 1.45 1.78 1.34 1.56 1.39 p Value 0.95 0.46 0.17 0.45 0.28 0.093 0.43 0.20 0.32 Lower limit of 0.518 0.660 0.815 0.653 0.740 0.909 0.647 0.785 0.725 95% CI Upper limit of 2.01 2.51 3.17 2.62 2.85 3.47 2.76 3.10 2.66 95% CI OR Quartile 4 1.29 1.51 1.78 1.31 1.66 1.52 1.22 1.77 1.29 p Value 0.53 0.31 0.13 0.52 0.23 0.28 0.65 0.19 0.50 Lower limit of 0.581 0.679 0.838 0.575 0.732 0.717 0.518 0.761 0.616 95% CI Upper limit of 2.88 3.35 3.79 2.98 3.75 3.20 2.86 4.10 2.71 95% CI

TABLE 30.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 381 371 330 378 330 375 Average 484 535 460 545 466 537 Stdev 385 578 384 569 400 555 p (t-test) 0.55 0.33 0.43 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 58 93 53 98 47 104 sCr only Median 347 376 330 378 324 386 Average 454 555 447 559 439 561 Stdev 380 581 379 579 382 573 p (t-test) 0.24 0.19 0.15 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 60 91 59 92 57 94 UO only Median 331 397 329 397 334 390 Average 481 555 477 557 491 531 Stdev 542 432 546 432 560 424 p (t-test) 0.39 0.35 0.63 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1880 4690 1880 4690 1880 n (Patient) 196 154 92 158 86 164 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.51 0.56 0.57 0.55 0.57 0.57 0.55 0.58 0.54 SE 0.048 0.047 0.049 0.049 0.047 0.048 0.050 0.047 0.048 p Value 0.82 0.21 0.16 0.34 0.15 0.13 0.36 0.077 0.36 nCohort Recovered 58 60 96 53 59 92 47 57 86 nCohort Non- 93 91 54 98 92 58 104 94 64 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity 75% 78% 78% 77% 78% 78% 77% 79% 77% Specificity 26% 30% 27% 28% 31% 27% 30% 32% 27% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity 49% 52% 57% 52% 52% 59% 52% 53% 55% Specificity 48% 52% 54% 53% 53% 55% 53% 54% 53% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity 25% 26% 30% 26% 27% 29% 25% 28% 28% Specificity 74% 77% 77% 75% 78% 77% 74% 79% 77% OR Quartile 2 1.06 1.52 1.30 1.29 1.58 1.29 1.41 1.71 1.19 p Value 0.88 0.27 0.51 0.51 0.23 0.51 0.38 0.16 0.65 Lower limit of 0.500 0.724 0.594 0.603 0.752 0.598 0.652 0.810 0.563 95% CI Upper limit of 2.25 3.20 2.85 2.75 3.32 2.79 3.07 3.60 2.52 95% CI OR Quartile 3 0.913 1.14 1.59 1.22 1.21 1.76 1.23 1.35 1.39 p Value 0.79 0.69 0.17 0.57 0.57 0.095 0.56 0.37 0.32 Lower limit of 0.474 0.595 0.813 0.622 0.628 0.906 0.616 0.700 0.725 95% CI Upper limit of 1.76 2.19 3.12 2.37 2.32 3.43 2.45 2.62 2.66 95% CI OR Quartile 4 0.942 1.18 1.42 1.05 1.32 1.40 0.972 1.43 1.29 p Value 0.88 0.67 0.37 0.89 0.48 0.37 0.94 0.37 0.50 Lower limit of 0.443 0.551 0.667 0.486 0.612 0.665 0.440 0.657 0.616 95% CI Upper limit of 2.00 2.51 3.01 2.28 2.85 2.96 2.15 3.13 2.71 95% CI

TABLE 30.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 364 386 348 392 348 392 Average 529 499 520 510 520 510 Stdev 611 368 610 376 610 376 p (t-test) 0.72 0.90 0.90 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1540 4690 1540 4690 1540 n (Patient) 81 70 80 71 80 71 sCr only Median 364 378 364 378 364 378 Average 456 563 455 561 455 561 Stdev 375 597 374 593 374 593 p (t-test) 0.20 0.21 0.21 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 67 84 65 86 65 86 UO only Median 328 428 328 428 328 428 Average 479 558 475 561 475 558 Stdev 555 405 553 413 559 408 p (t-test) 0.36 0.31 0.33 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1880 4690 1880 4690 1880 n (Patient) 95 55 93 57 91 59 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.53 0.56 0.59 0.54 0.56 0.59 0.54 0.56 0.59 SE 0.047 0.047 0.049 0.047 0.047 0.048 0.047 0.047 0.048 p Value 0.59 0.20 0.059 0.43 0.21 0.062 0.43 0.21 0.058 nCohort Recovered 81 67 95 80 65 93 80 65 91 nCohort Non- 70 84 55 71 86 57 71 86 59 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity 76% 79% 80% 76% 78% 79% 76% 78% 80% Specificity 26% 30% 28% 26% 29% 28% 26% 29% 29% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity 53% 52% 60% 54% 52% 60% 54% 52% 59% Specificity 52% 52% 56% 52% 52% 56% 52% 52% 56% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity 26% 27% 31% 27% 28% 32% 27% 28% 32% Specificity 75% 78% 78% 76% 78% 78% 76% 78% 79% OR Quartile 2 1.09 1.56 1.59 1.13 1.46 1.46 1.13 1.46 1.57 p Value 0.82 0.24 0.26 0.74 0.32 0.35 0.74 0.32 0.26 Lower limit of 0.521 0.746 0.716 0.540 0.696 0.666 0.540 0.696 0.718 95% CI Upper limit of 2.28 3.27 3.52 2.37 3.05 3.18 2.37 3.05 3.42 95% CI OR Quartile 3 1.21 1.20 1.89 1.27 1.20 1.87 1.27 1.20 1.86 p Value 0.56 0.57 0.064 0.46 0.57 0.066 0.46 0.57 0.067 Lower limit of 0.636 0.633 0.964 0.671 0.632 0.960 0.671 0.632 0.957 95% CI Upper limit of 2.29 2.29 3.72 2.41 2.29 3.66 2.41 2.29 3.61 95% CI OR Quartile 4 1.06 1.31 1.58 1.17 1.41 1.68 1.17 1.41 1.80 p Value 0.89 0.48 0.23 0.67 0.37 0.17 0.67 0.37 0.12 Lower limit of 0.506 0.618 0.745 0.562 0.662 0.799 0.562 0.662 0.855 95% CI Upper limit of 2.20 2.76 3.34 2.45 3.00 3.55 2.45 3.00 3.79 95% CI

Example 31. Use of C—C Motif Chemokine 24 for Evaluating Renal Status in Patients Admitted to the ICU: Recovery to RIFLE 0 and R from RIFLE I and F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 24 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “recovered” and a “non-recovered” population. “Recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-recovered” indicates those patients whose maximum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the recovery period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “non-recovered”.

The ability to distinguish the “recovered” and “non-recovered” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 31.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 12.9 40.4 12.2 40.4 12.5 39.7 Average 40.8 91.5 39.2 91.3 40.0 90.2 Stdev 87.7 166 88.2 165 89.4 164 p (t-test) 0.0035 0.0029 0.0044 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 670 1150 670 1150 670 1150 n (Patient) 107 209 103 213 100 216 sCr only Median 15.8 39.8 14.5 40.3 14.5 40.4 Average 51.4 88.2 51.4 87.7 51.2 87.7 Stdev 125 157 126 156 127 156 p (t-test) 0.029 0.031 0.031 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 125 190 123 192 122 193 UO only Median 20.4 51.1 18.4 51.1 18.4 50.6 Average 54.7 121 51.1 119 52.0 112 Stdev 117 193 117 184 120 178 p (t-test) 2.5E−4 9.3E−5 4.7E−4 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 221 93 207 107 197 117 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.65 0.63 0.68 0.66 0.64 0.71 0.66 0.64 0.69 SE 0.031 0.031 0.034 0.031 0.031 0.032 0.031 0.031 0.032 p Value 1.5E−6 2.5E−5 9.7E−8 1.7E−7 1.3E−5 8.3E−11 4.8E−7 7.2E−6 1.3E−9 nCohort 107 125 221 103 123 207 100 122 197 Recovered nCohort Non- 209 190 93 213 192 107 216 193 117 recovered Cutoff 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Quartile 2 Sensitivity 80% 81% 88% 81% 81% 90% 81% 81% 88% Specificity 36% 34% 31% 37% 35% 33% 37% 35% 33% Cutoff 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Quartile 3 Sensitivity 59% 59% 69% 59% 60% 71% 58% 60% 68% Specificity 67% 64% 58% 69% 65% 61% 68% 66% 61% Cutoff 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Quartile 4 Sensitivity 32% 32% 43% 32% 32% 43% 31% 32% 40% Specificity 88% 86% 82% 89% 85% 84% 89% 86% 84% OR Quartile 2 2.26 2.24 3.31 2.45 2.33 4.27 2.43 2.37 3.62 p Value 0.0023 0.0022 6.8E−4 8.2E−4 0.0014 3.6E−5 9.6E−4 0.0011 6.6E−5 Lower limit 1.34 1.34 1.66 1.45 1.39 2.15 1.44 1.41 1.92 of 95% CI Upper limit 3.81 3.76 6.61 4.15 3.91 8.50 4.12 3.99 6.82 of 95% CI OR Quartile 3 2.94 2.61 3.04 3.21 2.78 3.81 2.98 2.87 3.37 p Value 1.5E−5 5.5E−5 2.2E−5 4.5E−6 2.0E−5 1.8E−7 1.9E−5 1.2E−5 8.4E−7 Lower limit 1.80 1.64 1.82 1.95 1.74 2.31 1.80 1.79 2.08 of 95% CI Upper limit 4.80 4.16 5.08 5.29 4.44 6.30 4.90 4.60 5.46 of 95% CI OR Quartile 4 3.34 2.81 3.52 3.92 2.72 3.98 3.72 2.92 3.46 p Value 2.7E−4 5.3E−4 4.3E−6 1.0E−4 8.2E−4 4.0E−7 1.9E−4 4.1E−4 4.2E−6 Lower limit 1.74 1.57 2.06 1.97 1.51 2.33 1.87 1.61 2.04 of 95% CI Upper limit 6.39 5.04 6.02 7.81 4.88 6.78 7.40 5.30 5.88 of 95% CI

TABLE 31.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 16.9 43.2 14.5 43.2 14.5 42.2 Average 38.1 101 37.6 101 37.1 100.0 Stdev 73.1 178 73.5 177 73.5 176 p (t-test) 1.3E−4 1.3E−4 1.5E−4 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 670 1150 670 1150 670 1150 n (Patient) 135 181 133 183 129 187 sCr only Median 17.9 42.7 17.6 42.2 17.2 42.7 Average 46.2 99.1 46.2 98.8 46.0 98.6 Stdev 111 168 111 168 112 167 p (t-test) 0.0012 0.0014 0.0014 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 148 168 147 169 146 170 UO only Median 20.4 69.8 19.9 69.8 19.7 55.0 Average 53.3 127 51.1 127 51.6 117 Stdev 117 195 115 192 118 183 p (t-test) 4.9E−5 1.9E−5 1.5E−4 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 225 89 219 95 206 108 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.65 0.70 0.67 0.65 0.71 0.67 0.66 0.69 SE 0.030 0.030 0.034 0.030 0.030 0.033 0.030 0.030 0.032 p Value 1.2E−7 4.5E−7 2.9E−9 3.9E−8 3.6E−7 1.4E−10 3.7E−8 1.8E−7 2.7E−9 nCohort 135 148 225 133 147 219 129 146 206 Recovered nCohort Non- 181 168 89 183 169 95 187 170 108 recovered Cutoff Quartile 2 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Sensitivity 82% 83% 89% 82% 83% 89% 82% 83% 87% Specificity 34% 34% 31% 35% 34% 32% 35% 34% 32% Cutoff Quartile 3 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Sensitivity 61% 61% 72% 61% 62% 73% 60% 62% 69% Specificity 64% 63% 59% 65% 63% 60% 65% 64% 60% Cutoff Quartile 4 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Sensitivity 34% 35% 47% 34% 34% 47% 34% 35% 44% Specificity 87% 86% 84% 88% 86% 84% 88% 86% 84% OR Quartile 2 2.32 2.45 3.49 2.40 2.49 3.91 2.41 2.53 3.10 p Value 0.0015 8.5E−4 6.2E−4 9.3E−4 6.8E−4 1.8E−4 8.9E−4 5.3E−4 4.8E−4 Lower limit 1.38 1.45 1.71 1.43 1.47 1.91 1.43 1.50 1.64 of 95% CI Upper limit 3.89 4.14 7.15 4.04 4.21 7.99 4.05 4.28 5.83 of 95% CI OR Quartile 3 2.81 2.68 3.63 2.98 2.76 3.95 2.85 2.83 3.44 p Value 1.2E−5 2.2E−5 2.1E−6 4.0E−6 1.4E−5 3.0E−7 1.0E−5 8.1E−6 1.0E−6 Lower limit 1.77 1.70 2.13 1.87 1.75 2.34 1.79 1.79 2.09 of 95% CI Upper limit 4.46 4.23 6.19 4.75 4.35 6.68 4.54 4.48 5.64 of 95% CI OR Quartile 4 3.62 3.19 4.54 3.84 3.14 4.90 3.95 3.35 4.19 p Value 2.2E−5 5.0E−5 5.4E−8 1.3E−5 6.5E−5 1.0E−8 1.3E−5 3.0E−5 1.6E−7 Lower limit 2.00 1.82 2.63 2.10 1.79 2.84 2.13 1.90 2.45 of 95% CI Upper limit 6.55 5.59 7.83 7.03 5.49 8.44 7.33 5.91 7.16 of 95% CI

TABLE 31.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 16.9 44.6 16.9 44.6 15.9 44.6 Average 49.3 97.8 49.6 96.9 49.2 96.2 Stdev 117 166 118 166 119 164 p (t-test) 0.0031 0.0040 0.0042 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 153 163 151 165 147 169 sCr only Median 17.2 44.7 16.9 44.6 16.4 44.7 Average 47.8 102 47.8 101 47.6 101 Stdev 112 171 112 170 113 170 p (t-test) 0.0010 0.0011 0.0011 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 160 156 159 157 158 158 UO only Median 23.0 58.3 22.9 56.7 21.0 54.5 Average 59.8 110 59.8 109 60.6 102 Stdev 137 163 138 162 141 155 p (t-test) 0.0062 0.0068 0.019 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 223 91 222 92 211 103 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.67 0.68 0.66 0.67 0.68 0.67 0.67 0.66 SE 0.030 0.030 0.035 0.030 0.030 0.035 0.030 0.030 0.034 p Value 8.2E−8 3.1E−8 4.0E−7 1.1E−7 2.5E−8 3.1E−7 3.3E−8 1.2E−8 1.1E−6 nCohort 153 160 223 151 159 222 147 158 211 Recovered nCohort Non- 163 156 91 165 157 92 169 158 103 recovered Cutoff Quartile 2 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Sensitivity 83% 84% 86% 83% 84% 86% 83% 84% 85% Specificity 33% 34% 30% 34% 34% 30% 35% 34% 30% Cutoff Quartile 3 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Sensitivity 63% 63% 68% 62% 64% 68% 62% 64% 67% Specificity 63% 63% 57% 64% 64% 58% 64% 64% 58% Cutoff Quartile 4 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Sensitivity 36% 36% 45% 35% 36% 45% 35% 36% 42% Specificity 86% 86% 83% 86% 86% 83% 86% 86% 83% OR Quartile 2 2.41 2.67 2.52 2.50 2.72 2.57 2.68 2.76 2.55 p Value 0.0011 3.6E−4 0.0055 6.9E−4 2.8E−4 0.0046 2.7E−4 2.2E−4 0.0031 Lower limit 1.42 1.56 1.31 1.47 1.58 1.34 1.58 1.61 1.37 of 95% CI Upper limit 4.09 4.58 4.85 4.23 4.65 4.94 4.54 4.74 4.75 of 95% CI OR Quartile 3 2.90 2.97 2.88 2.90 3.06 2.96 2.91 3.14 2.84 p Value 5.1E−6 3.0E−6 5.6E−5 5.0E−6 1.8E−6 3.6E−5 4.9E−6 1.0E−6 3.5E−5 Lower limit 1.83 1.88 1.72 1.84 1.93 1.77 1.84 1.98 1.73 of 95% CI Upper limit 4.57 4.70 4.82 4.58 4.83 4.95 4.60 4.97 4.65 of 95% CI OR Quartile 4 3.47 3.34 3.99 3.36 3.28 3.89 3.41 3.49 3.48 p Value 1.4E−5 1.7E−5 5.2E−7 2.3E−5 2.3E−5 7.9E−7 2.3E−5 1.0E−5 4.1E−6 Lower limit 1.98 1.93 2.32 1.91 1.89 2.27 1.93 2.00 2.05 of 95% CI Upper limit 6.09 5.78 6.86 5.88 5.68 6.68 6.01 6.08 5.93 of 95% CI

TABLE 31.4 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 16.5 49.9 16.7 48.2 16.2 48.2 Average 53.9 97.2 54.5 95.6 54.2 94.9 Stdev 127 163 128 161 129 160 p (t-test) 0.0083 0.012 0.013 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 167 149 164 152 160 156 sCr only Median 16.9 49.9 16.7 49.5 16.5 49.9 Average 52.1 100 52.2 100 52.0 99.9 Stdev 123 167 123 166 124 166 p (t-test) 0.0033 0.0036 0.0036 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 171 145 170 146 169 147 UO only Median 20.7 64.0 21.0 58.3 21.0 54.7 Average 59.5 110 59.8 108 60.5 103 Stdev 138 161 138 161 140 156 p (t-test) 0.0057 0.0072 0.016 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 222 92 221 93 213 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.67 0.69 0.67 0.68 0.69 0.67 0.68 0.67 SE 0.030 0.030 0.034 0.030 0.030 0.034 0.030 0.030 0.034 p Value 8.1E−9 1.0E−8 1.3E−8 3.3E−8 8.2E−9 4.6E−8 1.1E−8 3.8E−9 5.5E−7 nCohort 167 171 222 164 170 221 160 169 213 Recovered nCohort Non- 149 145 92 152 146 93 156 147 101 recovered Cutoff Quartile 2 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Sensitivity 84% 84% 88% 84% 84% 87% 84% 84% 86% Specificity 33% 33% 31% 33% 33% 30% 34% 33% 31% Cutoff Quartile 3 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Sensitivity 66% 66% 71% 65% 66% 70% 65% 66% 67% Specificity 64% 63% 59% 64% 64% 58% 64% 64% 58% Cutoff Quartile 4 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Sensitivity 37% 37% 46% 36% 37% 45% 36% 37% 43% Specificity 86% 85% 83% 85% 85% 83% 86% 86% 83% OR Quartile 2 2.56 2.58 3.25 2.49 2.63 2.94 2.67 2.67 2.73 p Value 7.0E−4 6.9E−4 8.3E−4 8.8E−4 5.5E−4 0.0016 3.6E−4 4.4E−4 0.0020 Lower limit 1.49 1.49 1.63 1.46 1.52 1.50 1.56 1.54 1.45 of 95% CI Upper limit 4.40 4.47 6.49 4.27 4.55 5.74 4.58 4.62 5.15 of 95% CI OR Quartile 3 3.43 3.26 3.40 3.32 3.34 3.26 3.32 3.43 2.87 p Value 1.9E−7 5.6E−7 4.3E−6 3.4E−7 3.2E−7 7.8E−6 3.4E−7 1.8E−7 3.2E−5 Lower limit 2.16 2.05 2.02 2.10 2.10 1.94 2.09 2.16 1.75 of 95% CI Upper limit 5.45 5.17 5.73 5.27 5.31 5.46 5.26 5.46 4.72 of 95% CI OR Quartile 4 3.49 3.47 4.20 3.31 3.40 4.10 3.34 3.61 3.65 p Value 7.2E−6 6.9E−6 2.0E−7 1.7E−5 9.2E−6 3.1E−7 1.7E−5 4.0E−6 2.0E−6 Lower limit 2.02 2.02 2.44 1.92 1.98 2.39 1.93 2.09 2.14 of 95% CI Upper limit 6.02 5.96 7.22 5.70 5.85 7.03 5.78 6.24 6.21 of 95% CI

TABLE 31.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 19.7 49.9 20.1 48.2 19.9 49.2 Average 60.2 99.3 60.8 97.7 60.8 97.3 Stdev 141 153 141 152 142 152 p (t-test) 0.022 0.030 0.032 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 202 114 200 116 199 117 sCr only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 18.8 46.0 18.8 46.0 18.8 46.0 Average 53.6 103 53.6 103 53.6 103 Stdev 120 173 120 173 120 173 p (t-test) 0.0028 0.0028 0.0028 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 184 132 184 132 184 132 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 21.9 54.2 22.7 53.9 22.7 53.9 Average 61.1 103 61.4 102 61.7 101 Stdev 140 158 140 157 141 156 p (t-test) 0.019 0.023 0.028 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 216 98 215 99 213 101 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.66 0.67 0.66 0.66 0.67 0.66 0.66 0.67 SE 0.032 0.031 0.034 0.033 0.031 0.034 0.032 0.031 0.034 p Value 1.7E−7 4.6E−7 3.0E−7 8.0E−7 4.6E−7 8.5E−7 5.6E−7 4.6E−7 1.0E−6 nCohort Recovered 202 184 216 200 184 215 199 184 213 nCohort Non- 114 132 98 116 132 99 117 132 101 recovered Cutoff Quartile 2 4.17 4.17 4.13 4.17 4.17 4.13 4.17 4.17 4.13 Sensitivity 87% 84% 88% 86% 84% 87% 86% 84% 87% Specificity 32% 32% 31% 32% 32% 31% 32% 32% 31% Cutoff Quartile 3 29.6 29.6 28.8 29.6 29.6 28.8 29.6 29.6 28.8 Sensitivity 68% 65% 68% 66% 65% 68% 67% 65% 67% Specificity 60% 61% 58% 60% 61% 58% 60% 61% 58% Cutoff Quartile 4 74.9 74.9 74.6 74.9 74.9 74.6 74.9 74.9 74.6 Sensitivity 39% 39% 42% 39% 39% 41% 38% 39% 41% Specificity 83% 85% 82% 83% 85% 82% 83% 85% 82% OR Quartile 2 3.06 2.43 3.22 2.87 2.43 2.93 2.92 2.43 3.04 p Value 3.9E−4 0.0019 6.1E−4 6.4E−4 0.0019 0.0012 5.2E−4 0.0019 8.1E−4 Lower limit of 95% CI 1.65 1.39 1.65 1.57 1.39 1.53 1.60 1.39 1.59 Upper limit of 95% CI 5.68 4.26 6.29 5.27 4.26 5.62 5.36 4.26 5.83 OR Quartile 3 3.11 2.91 3.03 2.90 2.91 2.91 2.98 2.91 2.87 p Value 4.1E−6 6.7E−6 1.7E−5 1.2E−5 6.7E−6 2.9E−5 7.7E−6 6.7E−6 3.2E−5 Lower limit of 95% CI 1.92 1.83 1.83 1.80 1.83 1.76 1.85 1.83 1.75 Upper limit of 95% CI 5.04 4.63 5.01 4.68 4.63 4.80 4.80 4.63 4.72 OR Quartile 4 3.22 3.51 3.37 3.09 3.51 3.29 3.03 3.51 3.15 p Value 1.3E−5 4.0E−6 7.8E−6 2.5E−5 4.0E−6 1.1E−5 3.4E−5 4.0E−6 2.2E−5 Lower limit of 95% CI 1.90 2.06 1.98 1.83 2.06 1.93 1.80 2.06 1.85 Upper limit of 95% CI 5.45 5.98 5.74 5.23 5.98 5.60 5.12 5.98 5.35

TABLE 31.6 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 330 386 344 380 368 376 Average 417 562 419 560 429 553 Stdev 315 579 318 576 321 573 p (t-test) 0.10 0.11 0.17 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1640 4690 1640 4690 1640 4690 n (Patient) 49 102 48 103 46 105 sCr only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 327 386 327 386 330 380 Average 437 555 437 555 441 552 Stdev 379 569 379 569 382 567 p (t-test) 0.18 0.18 0.21 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 52 98 52 98 51 99 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 368 384 371 376 364 392 Average 505 514 485 549 487 540 Stdev 546 404 538 440 550 428 p (t-test) 0.92 0.47 0.53 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1880 4690 1880 4690 1880 n (Patient) 104 46 97 53 91 59 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.57 0.57 0.53 0.57 0.57 0.56 0.55 0.57 0.56 SE 0.049 0.048 0.052 0.049 0.048 0.050 0.050 0.049 0.048 p Value 0.15 0.13 0.54 0.16 0.13 0.25 0.29 0.16 0.25 nCohort Recovered 49 52 104 48 52 97 46 51 91 nCohort Non- 102 98 46 103 98 53 105 99 59 recovered Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 75% 77% 78% 76% 77% 79% 75% 77% 78% Specificity 27% 29% 27% 27% 29% 28% 26% 29% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 53% 53% 52% 52% 53% 51% 51% 53% 53% Specificity 55% 56% 51% 54% 56% 51% 52% 55% 52% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 29% 29% 30% 29% 29% 32% 29% 28% 31% Specificity 84% 81% 77% 83% 81% 78% 83% 80% 78% OR Quartile 2 1.11 1.32 1.33 1.16 1.32 1.47 1.07 1.38 1.34 p Value 0.79 0.47 0.50 0.71 0.47 0.34 0.86 0.41 0.46 Lower limit of 95% CI 0.511 0.618 0.582 0.531 0.618 0.663 0.485 0.643 0.621 Upper limit of 95% CI 2.42 2.83 3.02 2.53 2.83 3.27 2.37 2.95 2.89 OR Quartile 3 1.38 1.43 1.13 1.30 1.43 1.06 1.16 1.35 1.18 p Value 0.36 0.30 0.72 0.45 0.30 0.86 0.68 0.39 0.62 Lower limit of 95% CI 0.697 0.725 0.566 0.655 0.725 0.543 0.577 0.684 0.614 Upper limit of 95% CI 2.74 2.80 2.27 2.59 2.80 2.07 2.31 2.65 2.28 OR Quartile 4 2.14 1.68 1.46 2.05 1.68 1.71 1.90 1.62 1.56 p Value 0.087 0.21 0.34 0.10 0.21 0.16 0.15 0.25 0.24 Lower limit of 95% CI 0.895 0.742 0.671 0.861 0.742 0.805 0.794 0.714 0.741 Upper limit of 95% CI 5.09 3.80 3.17 4.90 3.80 3.63 4.54 3.66 3.28

TABLE 31.7 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 331 392 331 392 348 380 Average 453 561 453 561 461 553 Stdev 367 594 367 594 369 590 p (t-test) 0.20 0.20 0.28 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 64 87 64 87 62 89 sCr only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 330 394 330 394 331 392 Average 446 570 446 570 449 566 Stdev 368 599 368 599 370 596 p (t-test) 0.14 0.14 0.16 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 67 84 67 84 66 85 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 337 396 337 396 337 396 Average 493 543 487 554 496 529 Stdev 544 404 542 413 557 403 p (t-test) 0.58 0.45 0.70 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1700 n (Patient) 105 45 103 47 95 55 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.55 0.57 0.56 0.55 0.57 0.57 0.53 0.56 0.55 SE 0.047 0.047 0.052 0.047 0.047 0.051 0.048 0.047 0.049 p Value 0.29 0.15 0.23 0.29 0.15 0.16 0.46 0.18 0.34 nCohort Recovered 64 67 105 64 67 103 62 66 95 nCohort Non- 87 84 45 87 84 47 89 85 55 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity 75% 76% 80% 75% 76% 81% 74% 76% 76% Specificity 25% 27% 28% 25% 27% 28% 24% 27% 26% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity 54% 55% 56% 54% 55% 55% 53% 54% 55% Specificity 55% 55% 52% 55% 55% 52% 53% 55% 53% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity 29% 30% 33% 29% 30% 34% 28% 29% 31% Specificity 80% 81% 78% 80% 81% 79% 79% 80% 78% OR Quartile 2 0.985 1.18 1.53 0.985 1.18 1.65 0.916 1.22 1.15 p Value 0.97 0.67 0.33 0.97 0.67 0.24 0.82 0.60 0.72 Lower limit of 95% CI 0.468 0.562 0.655 0.468 0.562 0.711 0.432 0.583 0.533 Upper limit of 95% CI 2.07 2.46 3.56 2.07 2.46 3.85 1.94 2.55 2.50 OR Quartile 3 1.42 1.49 1.38 1.42 1.49 1.36 1.27 1.42 1.33 p Value 0.29 0.22 0.37 0.29 0.22 0.38 0.47 0.29 0.40 Lower limit of 95% CI 0.742 0.783 0.682 0.742 0.783 0.682 0.665 0.742 0.685 Upper limit of 95% CI 2.71 2.85 2.77 2.71 2.85 2.73 2.44 2.70 2.60 OR Quartile 4 1.58 1.76 1.78 1.58 1.76 1.90 1.47 1.70 1.58 p Value 0.24 0.15 0.14 0.24 0.15 0.10 0.32 0.17 0.23 Lower limit of 95% CI 0.736 0.819 0.823 0.736 0.819 0.884 0.684 0.790 0.745 Upper limit of 95% CI 3.40 3.78 3.86 3.40 3.78 4.09 3.17 3.65 3.34

TABLE 31.8 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “recovered” and “non-recovered” cohorts where recovery starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 332 386 348 380 348 380 Average 461 566 467 559 460 563 Stdev 373 612 372 611 363 610 p (t-test) 0.21 0.27 0.22 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 73 78 72 79 70 81 sCr only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 331 392 331 392 331 392 Average 448 580 448 580 448 580 Stdev 366 616 366 616 366 616 p (t-test) 0.11 0.11 0.11 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 74 77 74 77 74 77 UO only Recovery Period Duration (hr) 24 48 72 Non- Non- Non- Recovered recovered Recovered recovered Recovered recovered Cohort Cohort Cohort Cohort Cohort Cohort Median 332 396 330 397 334 394 Average 500 526 487 554 496 528 Stdev 550 385 576 401 555 405 p (t-test) 0.78 0.45 0.71 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1700 n (Patient) 105 45 103 47 96 54 Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.55 0.57 0.56 0.54 0.57 0.58 0.54 0.57 0.54 SE 0.047 0.047 0.052 0.047 0.047 0.051 0.047 0.047 0.049 p Value 0.29 0.13 0.28 0.42 0.13 0.11 0.41 0.13 0.37 nCohort Recovered 73 74 105 72 74 103 70 74 96 nCohort Non- 78 77 45 79 77 47 81 77 54 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity 77% 78% 80% 76% 78% 81% 75% 78% 76% Specificity 27% 28% 28% 26% 28% 28% 26% 28% 26% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity 54% 55% 58% 53% 55% 60% 53% 55% 56% Specificity 53% 54% 53% 53% 54% 54% 53% 54% 53% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity 27% 29% 29% 27% 29% 32% 27% 29% 30% Specificity 77% 78% 76% 76% 78% 78% 77% 78% 77% OR Quartile 2 1.26 1.40 1.53 1.13 1.40 1.65 1.06 1.40 1.11 p Value 0.54 0.37 0.33 0.74 0.37 0.24 0.89 0.37 0.78 Lower limit of 95% CI 0.602 0.668 0.655 0.543 0.668 0.711 0.506 0.668 0.513 Upper limit of 95% CI 2.63 2.93 3.56 2.36 2.93 3.85 2.20 2.93 2.40 OR Quartile 3 1.34 1.41 1.56 1.27 1.41 1.76 1.27 1.41 1.42 p Value 0.37 0.29 0.21 0.47 0..29 0.11 0.47 0.29 0.31 Lower limit of 95% CI 0.706 0.744 0.733 0.669 0.744 0.872 0.668 0.744 0.725 Upper limit of 95% CI 2.54 2.68 3.16 2.41 2.68 3.54 2.41 2.68 2.77 OR Quartile 4 1.21 1.45 1.30 1.17 1.45 1.63 1.26 1.45 1.42 p Value 0.61 0.33 0.51 0.67 0.33 0.21 0.54 0.33 0.37 Lower limit of 95% CI 0.580 0.690 0.593 0.560 0.690 0.756 0.599 0.690 0.667 Upper limit of 95% CI 2.54 3.05 2.85 2.45 3.05 3.52 2.64 3.05 3.01

Table 31.9: Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria.

TABLE 31.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Re- Non- Re- Non- Re- Non- covered recovered covered recovered covered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 348 392 364 386 364 386 Average 456 575 461 568 455 571 Stdev 368 621 368 620 359 619 p (t-test) 0.15 0.20 0.16 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n 76 75 75 76 73 78 (Patient) sCr only Median 330 397 330 397 330 397 Average 439 594 439 594 439 594 Stdev 363 623 363 623 363 623 p (t-test) 0.063 0.063 0.063 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n 77 74 77 74 77 74 (Patient) UO only Median 329 412 329 412 332 396 Average 487 550 487 550 493 532 Stdev 551 398 551 398 555 407 p (t-test) 0.47 0.47 0.65 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1700 n 100 50 100 50 95 55 (Patient) Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC  0.56  0.59  0.58  0.55  0.59  0.58  0.55  0.59  0.55 SE  0.047  0.046  0.050  0.047  0.046  0.050  0.047  0.046  0.049 p Value  0.22  0.052  0.11  0.33  0.052  0.11  0.32  0.052  0.31 nCohort Recovered  76  77 100  75  77 100  73  77  95 nCohort Non-  75  74  50  76  74  50  78  74  55 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity  77%  80%  80%  76%  80%  80%  76%  80%  76% Specificity  28%  30%  28%  27%  30%  28%  26%  30%  26% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity  53%  55%  60%  53%  55%  60%  53%  55%  56% Specificity  53%  55%  55%  52%  55%  55%  52%  55%  54% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity  28%  30%  30%  28%  30%  30%  28%  30%  29% Specificity  78%  79%  77%  77%  79%  77%  78%  79%  77% OR Quartile 2  1.30  1.68  1.56  1.17  1.68  1.56  1.09  1.68  1.15 p Value  0.48  0.18  0.29  0.67  0.18  0.29  0.81  0.18  0.72 Lower limit of 95% CI  0.623  0.793  0.686  0.561  0.793  0.686  0.524  0.793  0.533 Upper limit of 95% CI  2.73  3.54  3.53  2.45  3.54  3.53  2.28  3.54  2.50 OR Quartile 3  1.27  1.49  1.83  1.20  1.49  1.83  1.20  1.49  1.50 p Value  0.46  0.22  0.085  0.57  0.22  0.085  0.57  0.22  0.24 Lower limit of 95% CI  0.670  0.785  0.920  0.636  0.785  0.920  0.635  0.785  0.767 Upper limit of 95% CI  2.41  2.83  3.65  2.28  2.83  3.65  2.28  2.83  2.92 OR Quartile 4  1.35  1.61  1.43  1.30  1.61  1.43  1.40  1.61  1.36 p Value  0.43  0.21  0.35  0.48  0.21  0.35  0.37  0.21  0.42 Lower limit of 95% CI  0.645  0.768  0.669  0.623  0.768  0.669  0.666  0.768  0.642 Upper limit of 95% CI  2.82  3.39  3.08  2.73  3.39  3.08  2.94  3.39  2.89

TABLE 31.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “recovered” and “non-recovered” cohorts where recovery starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Recovery Period Duration (hr) 24 48 72 Re- Non- Re- Non- Re Non- covered recovered covered recovered covered recovered Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 332 397 334 396 332 397 Average 509 525 514 518 505 529 Stdev 590 377 591 378 589 386 p (t-test) 0.85 0.96 0.78 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1540 4690 1540 4690 1540 n 89 62 88 63 87 64 (Patient) sCr only Median 332 397 332 397 332 397 Average 451 586 451 586 451 586 Stdev 373 625 373 625 373 625 p (t-test) 0.11 0.11 0.11 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n 79 72 79 72 79 72 (Patient) UO only Median 326 448 326 448 326 448 Average 489 544 489 544 485 548 Stdev 565 368 565 368 564 379 p (t-test) 0.53 0.53 0.47 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1510 4690 1510 4690 1510 n 98 52 98 52 96 54 (Patient) Recovery Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC  0.56  0.58  0.59  0.55  0.58  0.59  0.56  0.58  0.59 SE  0.048  0.047  0.050  0.048  0.047  0.050  0.048  0.047  0.049 p Value  0.22  0.096  0.068  0.34  0.096  0.068  0.23  0.096  0.072 nCohort Recovered  89  79  98  88  79  98  87  79  96 nCohort Non-  62  72  52  63  72  52  64  72  54 recovered Cutoff Quartile 2 218 218 217 218 218 217 218 218 217 Sensitivity  77%  79%  81%  76%  79%  81%  77%  79%  80% Specificity  27%  29%  29%  26%  29%  29%  26%  29%  28% Cutoff Quartile 3 373 373 372 373 373 372 373 373 372 Sensitivity  56%  56%  62%  56%  56%  62%  56%  56%  61% Specificity  54%  54%  56%  53%  54%  56%  54%  54%  56% Cutoff Quartile 4 674 674 661 674 674 661 674 674 661 Sensitivity  27%  29%  31%  27%  29%  31%  28%  29%  31% Specificity  76%  78%  78%  76%  78%  78%  77%  78%  78% OR Quartile 2 1.27 1.56 1.68 1.13 1.56 1.68 1.17 1.56 1.53 p Value  0.54  0.24  0.21  0.75  0.24  0.21  0.67  0.24  0.30 Lower limit of 95% CI  0.594  0.739  0.742  0.535  0.739  0.742  0.555  0.739  0.689 Upper limit of 95% CI  2.70  3.30  3.80  2.40  3.30  3.80  2.48  3.30  3.40 OR Quartile 3  1.52  1.49  2.05  1.43  1.49  2.05  1.51  1.49  2.02 p Value  0.21  0.22  0.041  0.28  0.22  0.041  0.21  0.22  0.043 Lower limit of 95% CI  0.790  0.786  1.03  0.748  0.786  1.03  0.789  0.786  1.02 Upper limit of 95% CI  2.91  2.84  4.07  2.74  2.84  4.07  2.89  2.84  3.99 OR Quartile 4  1.22  1.50  1.54  1.18  1.50  1.54  1.31  1.50  1.64 p Value  0.59  0.28  0.27  0.66  0.28  0.27  0.47  0.28  0.20 Lower limit of 95% CI  0.582  0.717  0.721  0.562  0.717  0.721  0.626  0.717  0.774 Upper limit of 95% CI  2.57  3.14  3.27  2.47  3.14  3.27  2.75  3.14  3.48

Example 32. Use of C—C Motif Chemokine 24 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 24 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0), risk of injury (R), or injury (I) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 32.1 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 19.2 54.8 19.9 72.5 23.1 73.8 Average 53.8 112 53.2 123 61.3 112 Stdev 120 180 116 191 136 168 p (t-test) 7.2E−4 8.7E−5 0.0070 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1140 1150 n 204 112 220 96 235 81 (Patient) sCr only Median 19.4 50.8 20.7 54.5 23.1 54.5 Average 58.7 102 59.1 108 66.1 95.1 Stdev 140 153 137 162 153 121 p (t-test) 0.012 0.0062 0.12 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1150 1140 1150 1140 1150 679 n 207 108 222 93 234 81 (Patient) UO only Median 25.9 97.0 26.1 105 25.9 109 Average 66.2 129 66.0 135 65.9 138 Stdev 138 191 137 197 137 199 p (t-test) 0.011 0.0069 0.0050 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n 274 40 277 37 278 36 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC  0.69  0.68  0.72  0.72  0.69  0.73  0.69  0.68  0.74 SE  0.032  0.033  0.048  0.033  0.034  0.049  0.036  0.036  0.049 p Value 3.1E−9 4.3E−8 5.3E−6 4.3E−11 1.9E−8 2.8E−6 7.3E−8 1.1E−6 1.2E−6 nCohort Non- 204 207 274 220 222 277 235 234 278 persistent nCohort Persistent 112 108  40  96  93  37  81  81  36 Cutoff Quartile 2  4.17  4.15  4.13  4.17  4.15  4.13  4.17  4.15  4.13 Sensitivity  86%  86%  92%  90%  89%  95%  89%  89%  94% Specificity  31%  31%  28%  31%  31%  28%  30%  30%  28% Cutoff Quartile 3  29.6  29.3  28.8  29.6  29.3  28.8  29.6  29.3  28.8 Sensitivity  68%  67%  75%  71%  69%  76%  69%  68%  78% Specificity  60%  58%  54%  59%  58%  53%  57%  56%  54% Cutoff Quartile 4  74.9  74.4  74.6  74.9  74.4  74.6  74.9  74.4  74.6 Sensitivity  43%  43%  57%  47%  44%  59%  47%  44%  61% Specificity  85%  84%  80%  85%  83%  79%  83%  82%  79% OR Quartile 2  2.68  2.77  4.73  3.93  3.74  6.74  3.39  3.41  6.51 p Value  0.0014  0.0013  0.012 1.7E−4 3.0E−4  0.0099  0.0013  0.0013  0.011 Lower limit of 95% CI  1.46  1.49  1.42  1.92  1.83  1.58  1.61  1.62  1.53 Upper limit of 95% CI  4.92  5.16  15.8  8.03  7.65  28.7  7.17  7.21  27.8 OR Quartile 3  3.14  2.81  3.47  3.51  3.01  3.57  2.92  2.69  4.04 p Value 3.8E−6 3.0E−5  0.0012 1.8E−6 2.6E−5 0.0015 9.3E−5 2.7E−4 8.5E−4 Lower limit of 95% CI  1.93  1.73  1.63  2.09  1.80  1.62  1.71  1.58  1.78 Upper limit of 95% CI  5.10  4.58  7.38  5.88  5.02  7.84  5.00  4.59  9.18 OR Quartile 4  4.19  3.91  5.27  4.83  3.82  5.66  4.18  3.55  6.09 p Value 1.6E−7 5.2E−7 2.6E−6 1.3E−8 1.1E−6 2.2E−6 3.7E−7 6.0E−6 1.2E−6 Lower limit of 95% CI  2.45  2.30  2.64  2.81  2.23  2.76  2.41  2.05  2.93 Upper limit of 95% CI  7.15  6.67  10.5  8.30  6.54  11.6  7.26  6.15  12.7

TABLE 32.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Per- Non- Per- Non- Per- persistent sistent persistent sistent persistent sistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 16.9 54.6 18.0 63.7 20.4 59.3 Average 52.1 106 51.2 116 59.6 107 Stdev 123 169 119 179 139 157 p (t-test) 0.0012 1.5E−4 0.0077 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1140 1150 n 185 131 203 113 218 98 (Patient) sCr only Median 18.3 50.9 18.4 54.6 20.4 50.9 Average 59.3 95.6 58.3 102 66.3 89.6 Stdev 146 144 141 151 158 113 p (t-test) 0.031 0.011 0.19 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1150 1140 1150 1140 1150 679 n 191 124 205 110 217 98 (Patient) UO only Median 24.4 78.9 24.4 82.3 24.4 86.0 Average 62.8 124 62.5 129 62.3 131 Stdev 135 182 134 187 134 188 p (t-test) 0.0040 0.0021 0.0016 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n 255 59 259 55 260 54 (Patient) Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC  0.70  0.68  0.69  0.73  0.70  0.70  0.70  0.68  0.71 SE  0.030  0.031  0.041  0.031  0.032  0.042  0.033  0.034  0.042 p Value 2.8E−11 6.6E−9 1.9E−6 1.8E−13 1.7E−10 1.0E−6 1.4E−9 1.3E−7 5.6E−7 nCohort Non- 185 191 255 203 205 259 218 217 260 persistent nCohort Persistent 131 124  59 113 110  55  98  98  54 Cutoff Quartile 2  4.17  4.15  4.13  4.17  4.15  4.13  4.17  4.15  4.13 Sensitivity  86%  86%  90%  90%  90%  91%  90%  90%  91% Specificity  33%  32%  29%  33%  33%  29%  32%  32%  28% Cutoff Quartile 3  29.6  29.3  28.8  29.6  29.3  28.8  29.6  29.3  28.8 Sensitivity  68%  67%  71%  72%  70%  73%  70%  68%  74% Specificity  63%  61%  55%  62%  60%  55%  59%  58%  55% Cutoff Quartile 4  74.9  74.4  74.6  74.9  74.4  74.6  74.9  74.4  74.6 Sensitivity  40%  40%  51%  44%  42%  53%  45%  42%  54% Specificity  86%  84%  81%  86%  84%  81%  84%  82%  81% OR Quartile 2  3.09  3.03  3.54  4.67  4.47  4.00  4.08  4.10  3.90 p Value 1.6E−4 2.6E−4  0.0052 1.1E−5 2.0E−5  0.0046 1.1E−4 1.1E−4  0.0054 Lower limit of 95% CI  1.72  1.67  1.46  2.35  2.25  1.53  2.00  2.01  1.49 Upper limit of 95% CI  5.54  5.48  8.60  9.28  8.88  10.4  8.32  8.38  10.2 OR Quartile 3  3.56  3.13  3.01  4.14  3.57  3.24  3.45  2.99  3.49 p Value 1.4E−7 2.3E−6 4.5E−4 2.2E−8 4.6E−7 3.4E−4 2.0E−6 2.0E−5 1.9E−4 Lower limit of 95% CI  2.22  1.95  1.63  2.52  2.18  1.70  2.07  1.81  1.81 Upper limit of 95% CI  5.72  5.03  5.56  6.81  5.86  6.15  5.75  4.95  6.73 OR Quartile 4  4.16  3.51  4.35  4.76  3.75  4.66  4.26  3.39  4.87 p Value 2.6E−7 3.6E−6 1.4E−6 1.5E−8 1.1E−6 8.5E−7 1.3E−7 7.1E−6 5.0E−7 Lower limit of 95% CI  2.42  2.06  2.39  2.77  2.20  2.53  2.49  1.99  2.63 Upper limit of 95% CI  7.14  5.96  7.91  8.18  6.37  8.60  7.29  5.77  9.03

TABLE 32.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 18.2 50.6 18.3 54.6 20.7 50.9 Average 52.1 102 50.2 110 59.8 101 Stdev 123 167 118 175 141 153 p (t-test) 0.0026 3.1E−4 0.017 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1140 1150 n (Patient) 174 142 189 127 204 112 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 19.2 48.2 19.7 50.6 22.7 48.2 Average 58.8 93.6 59.1 96.8 68.0 84.3 Stdev 146 145 144 146 161 110 p (t-test) 0.036 0.026 0.35 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1150 1140 1150 1140 1150 679 n (Patient) 181 134 194 121 207 108 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 24.4 73.5 24.9 73.6 24.9 73.6 Average 62.2 117 62.0 122 62.0 122 Stdev 135 177 134 182 134 182 p (t-test) 0.0063 0.0035 0.0035 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 245 69 250 64 250 64 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.65 0.67 0.70 0.67 0.67 0.67 0.65 0.67 SE 0.031 0.032 0.039 0.031 0.032 0.040 0.033 0.033 0.040 p Value 4.2E−8 1.1E−6 1.3E−5 1.2E−10 5.1E−8 1.5E−5 1.2E−7 8.7E−6 1.5E−5 nCohort Non- 174 181 245 189 194 250 204 207 250 persistent nCohort Persistent 142 134 69 127 121 64 112 108 64 Cutoff Quartile 2 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Sensitivity 82% 82% 86% 85% 85% 86% 85% 85% 86% Specificity 30% 30% 28% 32% 31% 28% 30% 30% 28% Cutoff Quartile 3 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Sensitivity 64% 63% 68% 68% 66% 69% 67% 65% 69% Specificity 61% 60% 55% 62% 60% 55% 59% 57% 55% Cutoff Quartile 4 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Sensitivity 39% 38% 46% 42% 40% 47% 41% 39% 47% Specificity 86% 85% 81% 86% 84% 80% 84% 82% 80% OR Quartile 2 1.95 2.00 2.31 2.64 2.62 2.38 2.44 2.52 2.38 p Value 0.014 0.012 0.024 9.4E−4 0.0012 0.025 0.0034 0.0029 0.025 Lower limit of 1.15 1.16 1.12 1.49 1.46 1.11 1.34 1.37 1.11 95% CI Upper limit of 3.33 3.44 4.78 4.70 4.71 5.07 4.43 4.62 5.07 95% CI OR Quartile 3 2.85 2.57 2.62 3.41 2.90 2.67 2.96 2.49 2.67 p Value 7.8E−6 6.0E−5 8.3E−4 3.9E−7 1.0E−5 1.0E−3 1.1E−5 2.0E−4 1.0E−3 Lower limit of 1.80 1.62 1.49 2.12 1.81 1.49 1.82 1.54 1.49 95% CI Upper limit of 4.51 4.07 4.62 5.47 4.66 4.78 4.79 4.03 4.78 95% CI OR Quartile 4 3.95 3.36 3.64 4.49 3.46 3.62 3.61 2.92 3.62 p Value 8.7E−7 8.4E−6 8.8E−6 6.2E−8 4.4E−6 1.5E−5 2.0E−6 6.3E−5 1.5E−5 Lower limit of 2.29 1.97 2.06 2.61 2.04 2.02 2.13 1.73 2.02 95% CI Upper limit of 6.83 5.72 6.44 7.73 5.87 6.48 6.13 4.94 6.48 95% CI

TABLE 32.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 18.2 49.9 18.2 54.6 19.7 50.9 Average 52.4 100 50.3 110 59.0 101 Stdev 123 167 119 174 141 152 p (t-test) 0.0035 3.6E-4 0.015 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1140 1150 n (Patient) 172 144 188 128 200 116 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 18.4 48.2 18.4 50.9 20.4 49.9 Average 58.8 93.0 56.9 99.2 65.2 88.8 Stdev 146 144 142 149 160 116 p (t-test) 0.039 0.012 0.17 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1150 1140 1150 1140 1150 679 n (Patient) 179 136 191 124 203 112 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 23.8 71.6 24.1 73.6 24.1 73.6 Average 62.3 111 61.9 117 61.9 117 Stdev 137 169 135 175 135 175 p (t-test) 0.011 0.0053 0.0053 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 238 76 244 70 244 70 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.67 0.66 0.67 0.70 0.68 0.68 0.68 0.66 0.68 SE 0.031 0.031 0.037 0.031 0.031 0.038 0.032 0.033 0.038 p Value 6.9E−8 5.4E−7 3.6E−6 1.1E−10 6.6E−9 2.2E−6 2.2E−8 6.5E−7 2.2E−6 nCohort Non- 172 179 238 188 191 244 200 203 244 persistent nCohort Persistent 144 136 76 128 124 70 116 112 70 Cutoff Quartile 2 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Sensitivity 82% 82% 87% 85% 85% 87% 85% 86% 87% Specificity 31% 31% 29% 32% 32% 29% 31% 31% 29% Cutoff Quartile 3 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Sensitivity 64% 64% 68% 68% 67% 70% 67% 66% 70% Specificity 62% 60% 56% 62% 61% 56% 60% 59% 56% Cutoff Quartile 4 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Sensitivity 38% 38% 46% 41% 40% 47% 41% 40% 47% Specificity 86% 85% 82% 86% 85% 81% 84% 83% 81% OR Quartile 2 2.02 2.07 2.69 2.69 2.76 2.73 2.62 2.70 2.73 p Value 0.0098 0.0087 0.0071 7.6E−4 6.6E−4 0.0090 0.0015 0.0013 0.0090 Lower limit of 1.19 1.20 1.31 1.51 1.54 1.28 1.44 1.47 1.28 95% CI Upper limit of 3.45 3.56 5.55 4.78 4.96 5.79 4.75 4.95 5.79 95% CI OR Quartile 3 2.84 2.70 2.74 3.50 3.13 2.94 3.08 2.76 2.94 p Value 8.0E−6 2.4E−5 3.0E−4 2.3E−7 2.3E−6 2.1E−4 4.4E−6 3.5E−5 2.1E−4 Lower limit of 1.80 1.70 1.59 2.18 1.95 1.66 1.91 1.71 1.66 95% CI Upper limit of 4.49 4.28 4.74 5.62 5.03 5.20 4.98 4.46 5.20 95% CI OR Quartile 4 3.81 3.49 3.76 4.40 3.77 3.84 3.85 3.34 3.84 p Value 1.6E−6 5.0E−6 3.1E−6 8.9E−8 1.1E−6 3.5E−6 6.9E−7 7.5E−6 3.5E−6 Lower limit of 2.21 2.04 2.16 2.56 2.21 2.17 2.26 1.97 2.17 95% CI Upper limit of 6.58 5.96 6.57 7.58 6.44 6.78 6.55 5.66 6.78 95% CI

TABLE 32.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non- persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 18.2 49.9 18.3 54.5 19.7 50.9 Average 52.8 98.7 50.9 107 59.5 98.5 Stdev 125 165 120 172 143 150 p (t-test) 0.0053 7.7E-4 0.021 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1140 1150 n (Patient) 168 148 183 133 196 120 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 18.4 48.2 18.4 50.9 20.4 49.9 Average 59.4 91.3 57.4 97.1 65.8 86.9 Stdev 148 142 144 147 161 114 p (t-test) 0.054 0.018 0.22 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1150 1140 1150 1140 1150 679 n (Patient) 175 140 187 128 199 116 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 23.7 55.1 23.8 62.5 23.8 62.5 Average 62.5 108 62.3 110 62.3 110 Stdev 138 165 137 168 137 168 p (t-test) 0.017 0.012 0.012 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 233 81 236 78 236 78 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.66 0.65 0.67 0.69 0.67 0.67 0.68 0.65 0.67 SE 0.031 0.031 0.037 0.031 0.031 0.037 0.032 0.033 0.037 p Value 1.3E−7 1.6E−6 4.5E−6 6.4E−10 2.7E−8 2.4E−6 2.6E−8 2.2E−6 2.4E−6 nCohort Non- 168 175 233 183 187 236 196 199 236 persistent nCohort Persistent 148 140 81 133 128 78 120 116 78 Cutoff Quartile 2 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Sensitivity 82% 81% 86% 85% 84% 87% 86% 84% 87% Specificity 31% 30% 29% 32% 32% 29% 32% 31% 29% Cutoff Quartile 3 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Sensitivity 64% 64% 68% 67% 66% 69% 67% 66% 69% Specificity 62% 61% 56% 62% 61% 56% 60% 59% 56% Cutoff Quartile 4 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Sensitivity 38% 38% 44% 41% 40% 45% 41% 40% 45% Specificity 86% 85% 82% 86% 85% 81% 85% 83% 81% OR Quartile 2 2.01 1.90 2.62 2.69 2.49 2.81 2.80 2.41 2.81 p Value 0.0099 0.018 0.0066 6.4E−4 0.0017 0.0050 6.8E−4 0.0033 0.0050 Lower limit of 1.18 1.12 1.31 1.52 1.41 1.37 1.55 1.34 1.37 95% CI Upper limit of 3.41 3.25 5.26 4.74 4.39 5.78 5.08 4.32 5.78 95% CI OR Quartile 3 2.83 2.68 2.72 3.34 3.09 2.91 3.03 2.71 2.91 p Value 8.3E−6 2.5E−5 2.4E−4 4.6E−7 2.6E−6 1.3E−4 5.0E−6 4.0E−5 1.3E−4 Lower limit of 1.79 1.69 1.59 2.09 1.93 1.68 1.88 1.68 1.68 95% CI Upper limit of 4.47 4.24 4.63 5.34 4.94 5.01 4.87 4.36 5.01 95% CI OR Quartile 4 3.84 3.49 3.53 4.32 3.76 3.55 3.82 3.31 3.55 p Value 1.7E−6 5.4E−6 6.6E−6 1.5E−7 1.2E−6 7.2E−6 8.2E−7 8.8E−6 7.2E−6 Lower limit of 2.21 2.04 2.04 2.50 2.20 2.04 2.24 1.95 2.04 95% CI Upper limit of 6.66 5.98 6.12 7.46 6.42 6.18 6.50 5.60 6.18 95% CI

TABLE 32.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 372 376 348 396 373 356 Average 482 573 466 624 527 483 Stdev 382 682 376 722 557 359 p (t-test) 0.30 0.080 0.64 Min 46.8 63.9 46.8 66.1 46.8 66.1 Max 1880 4690 1880 4690 4690 1700 n (Patient) 96 55 104 47 111 40 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 368 384 364 392 372 356 Average 473 582 476 598 521 496 Stdev 380 679 385 717 557 381 p (t-test) 0.21 0.18 0.79 Min 46.8 63.9 46.8 63.9 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 94 56 103 47 108 42 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 364 392 364 392 350 394 Average 518 435 518 435 517 441 Stdev 531 260 531 260 529 265 p (t-test) 0.50 0.50 0.55 Min 46.8 66.1 46.8 66.1 46.8 66.1 Max 4690 1050 4690 1050 4690 1050 n (Patient) 131 19 131 19 132 18 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.52 0.54 0.51 0.57 0.55 0.51 0.52 0.52 0.51 SE 0.049 0.049 0.071 0.051 0.051 0.071 0.054 0.053 0.073 p Value 0.61 0.42 0.94 0.15 0.33 0.94 0.74 0.69 0.89 nCohort 96 94 131 104 103 131 111 108 132 Non-persistent nCohort Persistent 55 56 19 47 47 19 40 42 18 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 76% 77% 84% 83% 81% 84% 82% 81% 83% Specificity 26% 27% 27% 29% 28% 27% 28% 28% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 51% 52% 58% 55% 53% 58% 50% 50% 61% Specificity 50% 51% 51% 52% 51% 51% 50% 50% 52% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 27% 29% 16% 30% 28% 16% 22% 24% 17% Specificity 76% 77% 73% 77% 76% 73% 74% 74% 73% OR Quartile 2 1.14 1.20 1.94 1.98 1.65 1.94 1.83 1.63 1.80 p Value 0.74 0.65 0.31 0.13 0.24 0.31 0.20 0.27 0.37 Lower limit of 95% CI 0.526 0.554 0.534 0.827 0.711 0.534 0.732 0.680 0.492 Upper limit of 95% CI 2.46 2.59 7.08 4.72 3.85 7.08 4.56 3.93 6.61 OR Quartile 3 1.04 1.12 1.44 1.34 1.20 1.44 0.982 1.00 1.67 p Value 0.91 0.74 0.46 0.41 0.60 0.46 0.96 1.0 0.32 Lower limit of 95% CI 0.534 0.578 0.544 0.669 0.604 0.544 0.477 0.490 0.610 Upper limit of 95% CI 2.01 2.17 3.81 2.67 2.40 3.81 2.02 2.04 4.57 OR Quartile 4 1.19 1.31 0.514 1.41 1.19 0.514 0.821 0.893 0.554 p Value 0.65 0.48 0.31 0.38 0.66 0.31 0.65 0.79 0.37 Lower limit of 95% CI 0.559 0.618 0.141 0.652 0.546 0.141 0.349 0.389 0.151 Upper limit of 95% CI 2.54 2.77 1.87 3.07 2.61 1.87 1.93 2.05 2.03

TABLE 32.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 348 396 348 396 373 356 Average 451 605 448 626 514 518 Stdev 359 662 353 689 557 409 p (t-test) 0.068 0.037 0.96 Min 46.8 63.9 46.8 63.9 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 88 63 94 57 101 50 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 331 397 330 397 364 392 Average 442 616 437 639 497 547 Stdev 354 668 348 690 555 422 p (t-test) 0.040 0.019 0.58 Min 46.8 63.9 46.8 63.9 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 88 62 93 57 99 51 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 364 392 364 392 350 394 Average 508 505 508 505 507 512 Stdev 531 374 531 374 529 380 p (t-test) 0.97 0.97 0.96 Min 46.8 66.1 46.8 66.1 46.8 66.1 Max 4690 1510 4690 1510 4690 1510 n (Patient) 123 27 123 27 124 26 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.57 0.58 0.54 0.58 0.60 0.54 0.53 0.56 0.54 SE 0.048 0.048 0.062 0.048 0.048 0.062 0.050 0.050 0.063 p Value 0.12 0.077 0.57 0.091 0.041 0.57 0.53 0.22 0.53 nCohort Non-persistent 88 88 123 94 93 123 101 99 124 nCohort Persistent 63 62 27 57 57 27 50 51 26 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 79% 79% 81% 81% 81% 81% 80% 80% 81% Specificity 28% 28% 27% 29% 29% 27% 28% 28% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 54% 55% 56% 54% 56% 56% 50% 53% 58% Specificity 52% 53% 51% 52% 54% 51% 50% 52% 52% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 30% 31% 26% 32% 32% 26% 26% 27% 27% Specificity 78% 78% 75% 79% 78% 75% 75% 76% 75% OR Quartile 2 1.53 1.50 1.61 1.69 1.71 1.61 1.53 1.62 1.52 p Value 0.28 0.30 0.37 0.20 0.19 0.37 0.31 0.25 0.43 Lower limit of 95% CI 0.709 0.695 0.565 0.761 0.772 0.565 0.677 0.714 0.531 Upper limit of 95% CI 3.28 3.22 4.61 3.73 3.79 4.61 3.48 3.66 4.37 OR Quartile 3 1.28 1.39 1.31 1.30 1.49 1.31 0.980 1.20 1.45 p Value 0.45 0.32 0.52 0.44 0.24 0.52 0.95 0.61 0.39 Lower limit of 95% CI 0.671 0.725 0.568 0.671 0.767 0.568 0.498 0.608 0.619 Upper limit of 95% CI 2.46 2.67 3.03 2.51 2.89 3.03 1.93 2.35 3.42 OR Quartile 4 1.57 1.60 1.04 1.71 1.68 1.04 1.07 1.18 1.11 p Value 0.23 0.21 0.94 0.16 0.17 0.94 0.87 0.67 0.84 Lower limit of 95% CI 0.748 0.765 0.401 0.810 0.799 0.401 0.491 0.549 0.424 Upper limit of 95% CI 3.29 3.37 2.69 3.60 3.55 2.69 2.32 2.55 2.88

TABLE 32.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 332 394 331 396 348 392 Average 451 594 448 605 505 531 Stdev 361 644 358 655 572 397 p (t-test) 0.086 0.062 0.76 Min 46.8 50.8 46.8 50.8 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 83 68 86 65 94 57 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 331 394 331 394 332 392 Average 443 599 444 608 494 547 Stdev 357 647 353 662 567 413 p (t-test) 0.063 0.051 0.54 Min 46.8 50.8 46.8 50.8 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 82 68 86 64 93 57 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 332 396 332 396 331 397 Average 498 540 498 540 496 547 Stdev 538 383 538 383 536 386 p (t-test) 0.66 0.66 0.61 Min 46.8 66.1 46.8 66.1 46.8 66.1 Max 4690 1510 4690 1510 4690 1510 n (Patient) 115 35 115 35 116 34 Persistence Period Duration (hr) 24 48 72 sCr or sCr UO sCr or sCr UO sCr or sCr UO UO only only UO only only UO only only AUC 0.57 0.58 0.57 0.58 0.58 0.57 0.56 0.57 0.57 SE 0.047 0.047 0.057 0.047 0.047 0.057 0.049 0.049 0.057 p Value 0.15 0.11 0.22 0.11 0.11 0.22 0.23 0.15 0.20 nCohort Non-persistent 83 82 115 86 86 115 94 93 116 nCohort Persistent 68 68 35 65 64 35 57 57 34 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 78% 78% 83% 78% 78% 83% 81% 81% 82% Specificity 28% 28% 28% 28% 28% 28% 29% 29% 28% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 54% 54% 60% 55% 55% 60% 54% 54% 62% Specificity 53% 54% 53% 53% 53% 53% 52% 53% 53% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 31% 31% 31% 31% 30% 31% 28% 28% 32% Specificity 80% 79% 77% 79% 78% 77% 77% 76% 77% OR Quartile 2 1.35 1.38 1.86 1.41 1.38 1.86 1.69 1.71 1.78 p Value 0.43 0.40 0.21 0.37 0.40 0.21 0.20 0.19 0.25 Lower limit of 95% CI 0.641 0.651 0.707 0.662 0.648 0.707 0.761 0.772 0.673 Upper limit of 95% CI 2.86 2.91 4.91 3.00 2.95 4.91 3.73 3.79 4.70 OR Quartile 3 1.35 1.38 1.69 1.43 1.39 1.69 1.30 1.33 1.85 p Value 0.36 0.33 0.18 0.28 0.32 0.18 0.44 0.40 0.12 Lower limit of 95% CI 0.708 0.725 0.785 0.747 0.725 0.785 0.671 0.685 0.848 Upper limit of 95% CI 2.56 2.63 3.66 2.73 2.66 3.66 2.51 2.57 4.05 OR Quartile 4 1.73 1.71 1.49 1.68 1.49 1.49 1.28 1.26 1.58 p Value 0.15 0.16 0.35 0.17 0.29 0.35 0.52 0.55 0.29 Lower limit of 95% CI 0.827 0.814 0.649 0.801 0.711 0.649 0.604 0.595 0.682 Upper limit of 95% CI 3.64 3.59 3.44 3.52 3.12 3.44 2.70 2.67 3.64

TABLE 32.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by scrum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 332 394 330 397 330 397 Average 455 585 447 603 498 541 Stdev 363 638 360 650 576 398 p (t-test) 0.12 0.063 0.61 Min 50.8 46.8 46.8 50.8 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 81 70 85 66 91 60 sCr only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 330 396 329 397 329 397 Average 442 598 440 608 492 547 Stdev 359 642 356 652 576 403 p (t-test) 0.064 0.045 0.52 Min 46.8 50.8 46.8 50.8 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 81 69 84 66 90 60 UO only Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 332 396 331 397 330 397 Average 503 520 499 531 497 537 Stdev 549 369 548 365 546 368 p (t-test) 0.86 0.73 0.68 Min 50.8 46.8 46.8 66.1 46.8 66.1 Max 4690 1510 4690 1510 4690 1510 n (Patient) 109 41 110 40 111 39

TABLE 32.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non-persistent Persistent Non-persistent Persistent Non-persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 331 396 330 397 329 397 Average 451 584 448 593 496 542 Stdev 360 630 357 640 581 399 p (t-test) 0.11 0.082 0.59 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 4690 1700 n (Patient) 78 73 81 70 88 63 sCr only Median 330 396 329 397 329 397 Average 446 590 443 599 496 539 Stdev 361 636 358 646 581 401 p (t-test) 0.086 0.063 0.61 Min 46.8 50.8 46.8 50.8 46.8 63.9 Max 1880 4690 1880 4690 4690 1700 n (Patient) 79 71 82 68 88 62 UO only Median 334 394 334 394 332 396 Average 506 513 506 513 504 518 Stdev 551 366 551 366 549 370 p (t-test) 0.93 0.93 0.88 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1510 4690 1510 4690 1510 n (Patient) 108 42 108 42 109 41 Persistence Period Duration (hr) 24 48 72 sCr or sCr sCr or sCr sCr or sCr UO only UO only UO only UO only UO only UO only AUC 0.56 0.57 0.55 0.57 0.58 0.55 0.58 0.57 0.55 SE 0.047 0.047 0.053 0.047 0.053 0.048 0.048 0.048 0.054 p Value 0.19 0.13 0.35 0.14 0.095 0.35 0.11 0.12 0.33 nCohort 78 79 108 81 822 108 88 88 109 Non-persistent nCohort Persistent 73 71 42 70 68 42 63 62 41 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 77% 77% 81% 77% 78% 81% 81% 81% 80% Specificity 27% 28% 28% 27% 28% 28% 30% 30% 28% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 55% 55% 57% 56% 56% 57% 57% 56% 59% Specificity 54% 54% 53% 54% 55% 53% 55% 55% 53% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 30% 30% 29% 30% 29% 29% 29% 27% 29% Specificity 79% 78% 76% 79% 78% 76% 77% 76% 76% OR Quartile 2 1.21 1.33 1.63 1.26 1.38 1.63 1.78 1.75 1.57 p Value 0.61 0.46 0.27 0.54 0.40 0.27 0.15 0.16 0.32 Lower limit of 0.580 0.631 0.680 0.599 0.651 0.680 0.819 0.802 0.650 95% CI Upper limit of 2.54 2.79 3.93 2.64 2.91 3.93 3.88 3.81 3.77 95% CI OR Quartile 3 1.41 1.46 1.49 1.50 1.54 1.49 1.60 1.56 1.61 p Value 0.29 0.25 0.28 0.22 0.19 0.28 0.16 0.19 0.20 Lower limit of 0.745 0.765 0.726 0.786 0.807 0.726 0.834 0.809 0.777 95% CI Upper limit of 2.68 2.77 3.06 2.85 2.94 3.06 3.07 2.99 3.32 95% CI OR Quartile 4 1.67 1.53 1.26 1.61 1.48 1.26 1.36 1.21 1.32 p Value 0.18 0.26 0.57 0.21 0.30 0.57 0.42 0.62 0.50 Lower limit of 0.795 0.731 0.566 0.770 0.708 0.566 0.649 0.574 0.591 95% CI Upper limit of 3.51 3.21 2.81 3.38 3.10 2.81 2.85 2.53 2.95 95% CI

Example 33. Use of C—C Motif Chemokine 24 for Evaluating Renal Status in Patients Admitted to the ICU: Persistent at RIFLE I or F

Patients from the intensive care unit (ICU) with RIFLE stage of injury (I) or failure (F) are enrolled in the following study. EDTA anti-coagulated blood samples (10 mL) and urine samples (50 mL) are collected from each patient at enrollment, and at every 12 hours up to day 3, and then every 24 hours thereafter up to day 7 while the subject is hospitalized. C—C motif chemokine 24 is measured in the enrollment samples by standard immunoassay methods using commercially available assay reagents.

Kidney status is assessed by RIFLE criteria based on serum creatinine, urine output, or both serum creatinine and urine output. Two cohorts are defined to represent a “persistent” and a “non-persistent” population. “Persistent” indicates those patients whose minimum RIFLE stage during a period of 24, 48 or 72 hours is injury (I) or failure (F) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. “Non-persistent” indicates those patients who are not persistent at injury (I) or failure (F) and whose minimum RIFLE stage during a period of 24, 48 or 72 hours is non-injury (RIFLE 0) or risk of injury (R) where the persistence period can start from the time of sample collection to 24, 48, 72, 96 or 168 hours after sample collection. If a patient dies after injury (I) or failure (F) or is placed on renal replacement therapy (RRT) at any time from sample collection to 24, 48, 72, 96 or 168 hours after sample collection, the patient is considered “persistent”.

The ability to distinguish the “persistent” and “non-persistent” cohorts is determined using receiver operating characteristic (ROC) analysis.

TABLE 33.1 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median  15.2  39.8  19.3  42.2  19.8  44.7 Average  54.0  86.4  51.3  97.0  52.6  102 Stdev  132  153  117  168  115  175 p (t-test)   0.056   0.0054   0.0026 Min   0.00127   0.00127   0.00127   0.00127   0.00127   0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient)  118  198  157  159  178  138 sCr only Median  17.6  38.9  19.3  42.0  20.4  46.0 Average  54.6  86.7  50.8  97.3  53.1  102 Stdev  127  157  116  169  114  178 p (t-test)   0.054   0.0046   0.0033 Min   0.00127   0.00127   0.00127   0.00127   0.00127   0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient)  129  186  161  154  183  132 UO only Median  22.9  73.9  24.4  89.6  25.6  97.6 Average  57.8  127  59.3  138  60.8  142 Stdev  131  181  130  191  131  197 p (t-test) 3.2E−4 1.7E−4 2.6E−4 Min   0.00127   0.00127   0.00127   0.00127   0.00127   0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient)  240  74  255  59  262  52 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.63   0.62   0.70   0.64   0.64   0.71   0.65   0.65   0.71 SE   0.032   0.032   0.037   0.031   0.031   0.040   0.031   0.032   0.043 p Value 5.0E−5 1.9E−4 4.1E−8 1.2E−5 6.5E−6 1.2E−7 1.1E−6 4.2E−6 6.4E−7 nCohort  118  129  240  157  161  255  178  183  262 Non-persistent nCohort Persistent  198  186  74  159  154  59  138  132  52 Cutoff Quartile 2   4.17   4.15   4.13   4.17   4.15   4.13   4.17   4.15   4.13 Sensitivity 80% 81% 89% 81% 82% 90% 83% 83% 90% Specificity 34% 33% 30% 31% 32% 29% 31% 31% 28% Cutoff Quartile 3  29.6  29.3  28.8  29.6  29.3  28.8  29.6  29.3  28.8 Sensitivity 58% 59% 72% 60% 60% 73% 62% 62% 71% Specificity 64% 62% 57% 60% 60% 55% 60% 58% 54% Cutoff Quartile 4  74.9  74.4  74.6  74.9  74.4  74.6  74.9  74.4  74.6 Sensitivity 31% 32% 49% 35% 35% 54% 37% 36% 56% Specificity 86% 84% 82% 85% 84% 82% 84% 83% 81% OR Quartile 2   2.09   2.08   3.47   1.95   2.09   3.54   2.30   2.26   3.70 p Value   0.0052   0.0053   0.0019   0.012   0.0063   0.0052   0.0030   0.0039   0.0076 Upper limit of   1.25   1.24   1.58   1.16   1.23   1.46   1.33   1.30   1.42 95% CI Lower limit   3.51   3.49   7.59   3.29   3.53   8.60   3.97   3.94   9.67 of 95% CI OR Quartile 3   2.42   2.31   3.30   2.21   2.25   3.32   2.43   2.31   2.92 p Value 2.3E−4 3.6E−4 3.6E−5 5.3E−4 4.2E−4 1.6E−4 1.3E−4 3.5E−4   0.0012 Upper limit of   1.51   1.46   1.87   1.41   1.43   1.78   1.54   1.46   1.53 95% CI Lower limit of   3.86   3.66   5.81   3.47   3.53   6.21   3.84   3.65   5.58 95% CI OR Quartile 4   2.71   2.53   4.34   2.93   2.94   5.25   3.14   2.80   5.35 p Value   0.0010   0.0013 3.2E−7 1.1E−4 9.1E−5 6.9E−8 2.4E−5 1.2E−4 1.7E−7 Upper limit of   1.49   1.43   2.47   1.70   1.71   2.87   1.85   1.66   2.85 95% CI Lower limit of   4.91   4.47   7.62   5.05   5.04   9.58   5.34   4.73  10.0 95% CI

TABLE 33.2 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median  14.3  40.4  17.9  42.7  17.9  44.7 Average  47.1  89.0  47.9  97.0  49.1  101 Stdev  125  155  115  166  114  171 p (t-test)   0.015   0.0028   0.0014 Min   0.00127   0.00127   0.00127   0.00127   0.00127   0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient)  111  205  146  170  164  152 sCr only Median  15.8  39.7  18.3  42.7  17.9  44.7 Average  48.6  90.0  45.9  99.7  47.6  103 Stdev  119  159  109  170  110  174 p (t-test)   0.014 9.9E−4 6.6E−4 Min   0.00127   0.00127   0.00127   0.00127   0.00127   0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient)  125  190  153  162  168  147 UO only Median  19.9  55.1  23.0  74.0  23.8  74.7 Average  57.2  114  59.1  121  60.7  121 Stdev  135  165  134  173  135  176 p (t-test)   0.0015   0.0013   0.0023 Min   0.00127   0.00127   0.00127   0.00127   0.00127   0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient)  221  93  237  77  244  70 Persistence Period Duration (hr) 24 48 72 sCr or sCr or sCr or UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC   0.65   0.63   0.70   0.65   0.66   0.70   0.67   0.67   0.69 SE   0.031   0.031   0.034   0.031   0.031   0.037   0.030   0.031   0.038 p Value 1.6E−6 2.1E−5 4.1E−9 6.2E−7 3.8E−7 6.0E−8 5.1E−8 3.5E−8 4.0E−7 nCohort Non-persistent  111  125  221  146  153  237  164  168  244 nCohort Persistent  205  190  93  170  162  77  152  147  70 Cutoff Quartile 2   4.17   4.15   4.13   4.17   4.15   4.13   4.17   4.15   4.13 Sensitivity 80% 81% 88% 82% 83% 88% 84% 84% 89% Specificity 35% 34% 31% 33% 33% 30% 33% 33% 29% Cutoff Quartile 3  29.6  29.3  28.8  29.6  29.3  28.8  29.6  29.3  28.8 Sensitivity 59% 59% 72% 61% 61% 73% 63% 64% 71% Specificity 67% 64% 59% 62% 61% 57% 62% 62% 56% Cutoff Quartile 4  74.9  74.4  74.6  74.9  74.4  74.6  74.9  74.4  74.6 Sensitivity 32% 32% 46% 35% 35% 49% 37% 37% 50% Specificity 88% 86% 84% 86% 86% 83% 86% 85% 82% OR Quartile 2   2.23   2.24   3.31   2.20   2.39   3.17   2.49   2.70   3.18 p Value   0.0025   0.0022 6.8E−4   0.0030   0.0012   0.0026 8.8E−4 4.0E−4   0.0039 Lower limit of 95% CI   1.33   1.34   1.66   1.31   1.41   1.50   1.46   1.56   1.45 Upper limit of 95% CI   3.76   3.76   6.61   3.70   4.06   6.70   4.27   4.67   6.98 OR Quartile 3   2.88   2.61   3.75   2.54   2.50   3.59   2.82   2.88   3.20 p Value 1.8E−5 5.5E−5 8.6E−7 5.7E−5 7.3E−5 8.8E−6 8.5E−6 6.1E−6 7.7E−5 Lower limit of 95% CI   1.78   1.64   2.22   1.61   1.59   2.04   1.79   1.82   1.80 Upper limit of 95% CI   4.67   4.16   6.35   4.01   3.94   6.31   4.45   4.56   5.70 OR Quartile 4   3.58   2.81   4.42   3.35   3.23   4.66   3.58   3.32   4.55 p Value 1.2E−4 5.3E−4 7.7E−8 3.0E−5 3.4E−5 7.0E−8 5.7E−6 1.4E−5 2.0E−7 Lower limit of 95% CI   1.87   1.57   2.57   1.90   1.86   2.66   2.06   1.93   2.57 Upper limit of 95% CI   6.85   5.04   7.60   5.91   5.63   8.15   6.20   5.71   8.05

TABLE 33.3 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 72 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 13.6 40.3 16.9 43.2 16.9 44.6 Average 47.4 88.7 47.8 96.2 49.6 98.7 Stdev 125 155 116 164 116 168 p (t-test) 0.017 0.0032 0.0027 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 110 206 143 173 157 159 sCr only Median 14.5 39.7 17.2 43.2 17.2 44.6 Average 48.7 89.5 45.9 98.8 48.0 101 Stdev 120 159 110 169 111 172 p (t-test) 0.015 0.0012 0.0013 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 123 192 150 165 162 153 UO only Median 20.3 54.7 22.9 64.0 23.7 69.8 Average 58.5 106 60.1 109 61.7 109 Stdev 138 159 138 163 138 164 p (t-test) 0.0064 0.0069 0.012 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 211 103 224 90 231 83 Persistence Period Duration (hr) 24 48 72 sCr or sCr sCr or sCr sCr or sCr UO only UO only UO only UO only UO only UO only AUC 0.65 0.64 0.68 0.66 0.66 0.68 0.66 0.66 0.67 SE 0.031 0.031 0.033 0.030 0.030 0.035 0.030 0.030 0.036 p Value 1.9E−6 1.4E−5 9.0E−8 2.5E−7 1.7E−7 2.3E−7 6.2E−8 6.7E−8 1.6E−6 nCohort Non-persistent 110 123 211 143 150 224 157 162 231 nCohort Persistent 206 192 103 173 165 90 159 153 83 Cutoff Quartile 2 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Sensitivity 81% 81% 85% 82% 83% 86% 83% 84% 86% Specificity 35% 35% 30% 34% 34% 29% 33% 33% 29% Cutoff Quartile 3 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Sensitivity 59% 59% 69% 61% 61% 70% 63% 63% 69% Specificity 66% 64% 59 63% 62% 58% 63% 62% 57% Cutoff Quartile 4 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Sensitivity 32% 32% 43% 35% 35% 46% 36% 36% 46% Specificity 88% 86% 83% 87% 86% 83% 86% 85% 82% OR Quartile 2 2.28 2.33 2.55 2.31 2.52 2.47 2.42 2.56 2.42 p Value 0.0019 0.0014 0.0031 0.0016 6.1E−4 0.0066 0.0011 6.3E−4 0.010 Lower limit of 95% CI 1.35 1.39 1.37 1.37 1.49 1.29 1.42 1.49 1.23 Upper limit of 95% CI 3.84 3.91 4.75 3.90 4.28 4.76 4.12 4.39 4.75 OR Quartile 3 2.81 2.62 3.22 2.62 2.57 3.23 2.89 2.87 2.87 p Value 2.8E−5 5.3E−5 4.3E−6 3.5E−5 4.6E−5 1.1E−5 5.1E−6 6.3E−6 1.0E−4 Lower limit of 95% CI 1.73 1.64 1.96 1.66 1.63 1.91 1.83 1.82 1.69 Upper limit of 95% CI 4.55 4.19 5.31 4.14 4.06 5.44 4.57 4.53 4.89 OR Quartile 4 3.52 2.97 3.75 3.47 3.33 4.10 3.43 3.23 3.91 p Value 1.4E−4 3.3E−4 1.2E−6 2.3E−5 2.6E−5 3.4E−7 1.4E−5 2.5E−5 1.1E−6 Lower limit of 95% CI 1.84 1.64 2.20 1.95 1.90 2.38 1.97 1.87 2.26 Upper limit of 95% CI 6.73 5.39 6.39 6.16 5.84 7.04 5.97 5.57 6.77

TABLE 33.4 Comparison of marker levels and the area under the ROC curve (AUC in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 96 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr UO Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 14.3 40.2 16.4 42.7 15.9 44.6 Average 47.8 88.2 47.9 95.9 49.6 98.1 Stdev 126 155 117 164 117 167 p (t-test) 0.019 0.0036 0.0032 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 109 207 142 174 155 161 sCr only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 14.5 39.7 16.9 42.7 16.9 44.6 Average 48.7 89.5 45.9 98.4 48.1 100 Stdev 120 159 111 168 112 171 p (t-test) 0.015 0.0014 0.0014 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 123 192 149 166 161 154 UO only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 19.9 54.5 20.4 55.1 22.7 55.1 Average 58.7 104 60.2 106 62.0 105 Stdev 139 157 139 159 140 159 p (t-test) 0.0091 0.010 0.019 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 207 107 219 95 225 89 Persistence Period Duration (hr) 24 48 72 sCr sCr sCr UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.64 0.67 0.66 0.66 0.68 0.67 0.67 0.68 SE 0.031 0.031 0.033 0.030 0.030 0.034 0.030 0.030 0.035 p Value 4.8E−6 1.4E−5 1.4E−7 2.2E−7 1.5E−7 1.1E−7 3.4E−8 5.9E−8 5.9E−7 nCohort Non- 109 123 207 142 149 219 155 161 225 persistent nCohort Persistent 207 192 107 174 166 95 161 154 89 Cutoff Quartile 2 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Sensitivity 80% 81% 85% 82% 83% 86% 83% 84% 87% Specificity 35% 35% 30% 34% 34% 30% 34% 34% 30% Cutoff Quartile 3 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Sensitivity 58% 59% 68% 61% 61% 69% 63% 64% 69% Specificity 66% 64% 59% 63% 62% 58% 64% 63% 57% Cutoff Quartile 4 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Sensitivity 32% 32% 42% 34% 35% 44% 35% 36% 44% Specificity 88% 86% 84% 87% 86% 83% 86% 85% 82% OR Quartile 2 2.17 2.33 2.49 2.36 2.56 2.72 2.51 2.60 2.72 p Value 0.0037 0.0014 0.0033 0.0013 4.8E−4 0.0026 7.0E−4 5.0E−4 0.0035 Lower limit of 95% CI 1.29 1.39 1.35 1.40 1.51 1.42 1.47 1.52 1.39 Upper limit of 95% CI 3.65 3.91 4.57 3.97 4.35 5.22 4.26 4.46 5.33 OR Quartile 3 2.74 2.62 3.14 2.70 2.65 3.20 3.06 2.95 2.93 p Value 4.4E−5 5.3E−5 5.2E−6 2.1E−5 2.8E−5 8.7E−6 1.8E−6 3.8E−6 5.1E−5 Lower limit of 95% CI 1.69 1.64 1.92 1.71 1.68 1.92 1.93 1.86 1.74 Upper limit of 95% CI 4.44 4.19 5.14 4.26 4.17 5.35 4.83 4.66 4.92 OR Quartile 4 3.46 2.97 3.69 3.41 3.27 3.90 3.31 3.17 3.61 p Value 1.8E−4 3.3E−4 1.5E−6 3.0E−5 3.4E−5 7.0E−7 2.3E−5 3.3E−5 3.3E−6 Lower limit of 95% CI 1.81 1.64 2.17 1.92 1.87 2.28 1.90 1.84 2.10 Upper limit of 95% CI 6.61 5.39 6.28 6.06 5.74 6.67 5.77 5.47 6.19

TABLE 33.5 Comparison of marker levels and the area under the ROC curve (AUC) in urine samples for the “persistent” and “non-persistent” cohorts where persistence starts within 168 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 14.3 39.2 14.5 43.2 15.8 44.6 Average 48.8 87.0 48.1 94.4 50.2 95.8 Stdev 128 153 119 162 119 164 p (t-test) 0.029 0.0052 0.0055 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 105 211 137 179 149 167 sCr only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 14.5 39.2 15.8 42.7 15.9 44.6 Average 49.2 88.6 46.1 97.0 48.3 98.7 Stdev 122 158 112 166 113 169 p (t-test) 0.020 0.0020 0.0021 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1070 1150 1070 1150 1070 1150 n (Patient) 120 195 145 170 157 158 UO only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 19.9 50.6 20.4 54.5 21.9 52.8 Average 59.5 100 61.3 101 63.2 98.3 Stdev 141 154 142 154 143 154 p (t-test) 0.018 0.026 0.050 Min 0.00127 0.00127 0.00127 0.00127 0.00127 0.00127 Max 1140 1150 1140 1150 1140 1150 n (Patient) 201 113 211 103 216 98 24 48 72 sCr sCr sCr UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.64 0.64 0.67 0.66 0.66 0.67 0.67 0.67 0.66 SE 0.032 0.031 0.033 0.030 0.030 0.034 0.030 0.030 0.034 p Value 8.0E−6 1.6E−5 2.9E−7 8.7E−8 9.6E−8 3.2E−7 2.4E−8 3.9E−8 1.7E−6 nCohort Non- 105 120 201 137 145 211 149 157 216 persistent nCohort Persistent 211 195 113 179 170 103 167 158 98 Cutoff Quartile 2 4.17 4.15 4.13 4.17 4.15 4.13 4.17 4.15 4.13 Sensitivity 81% 82% 85% 83% 84% 86% 84% 84% 87% Specificity 36% 36% 31% 35% 35% 31% 35% 34% 31% Cutoff Quartile 3 29.6 29.3 28.8 29.6 29.3 28.8 29.6 29.3 28.8 Sensitivity 58% 59% 67% 61% 61% 68% 63% 63% 67% Specificity 66% 64% 60% 64% 63% 59% 64% 63% 58% Cutoff Quartile 4 74.9 74.4 74.6 74.9 74.4 74.6 74.9 74.4 74.6 Sensitivity 31% 32% 41% 34% 35% 42% 35% 35% 41% Specificity 88% 86% 84% 87% 86% 83% 86% 85% 82% OR Quartile 2 2.35 2.47 2.52 2.57 2.75 2.83 2.78 2.79 2.88 p Value 0.0014 6.6E−4 0.0024 3.9E−4 1.8E−4 0.0013 1.7E−4 1.9E−4 0.0015 Lower limit of 1.39 1.47 1.39 1.53 1.62 1.50 1.63 1.63 1.50 95% CI Upper limit of 3.97 4.15 4.57 4.34 4.67 5.34 4.73 4.78 5.52 95% CI OR Quartile 3 2.63 2.57 3.04 2.80 2.66 3.02 3.07 2.94 2.83 p Value 1.0E−4 8.0E−5 6.5E−6 1.2E−5 2.8E−5 1.3E−5 1.7E−6 3.8E−6 4.7E−5 Lower limit of 1.61 1.61 1.88 1.76 1.68 1.84 1.94 1.86 1.72 95% CI Upper limit of 4.28 4.12 4.94 4.43 4.19 4.97 4.86 4.65 4.68 95% CI OR Quartile 4 3.22 2.82 3.50 3.42 3.32 3.48 3.24 3.20 3.13 p Value 4.2E−4 6.3E−4 3.6E−6 3.7E−5 3.4E−5 4.1E−6 3.9E−5 3.4E−5 2.6E−5 Lower limit of 1.68 1.56 2.06 1.91 1.88 2.05 1.85 1.85 1.84 95% CI Upper limit of 6.17 5.12 5.93 6.13 5.86 5.93 5.68 5.54 5.33 95% CI

TABLE 33.6 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 24 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 347 380 364 378 373 355 Average 446 549 470 559 487 552 Stdev 374 566 370 620 381 644 p (t-test) 0.24 0.29 0.44 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 50 101 75 76 85 66 sCr only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 327 386 371 374 376 332 Average 437 555 481 545 491 544 Stdev 379 569 387 610 393 642 p (t-test) 0.18 0.45 0.54 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 52 98 73 77 86 64 UO only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 334 397 331 412 330 428 Average 504 520 504 524 509 503 Stdev 545 370 537 354 539 287 p (t-test) 0.87 0.84 0.96 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1050 n (Patient) 112 38 120 30 125 25 Persistence Period Duration (hr) 24 48 72 sCr sCr sCr UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.55 0.57 0.55 0.53 0.52 0.57 0.51 0.51 0.57 SE 0.049 0.048 0.055 0.047 0.047 0.060 0.048 0.048 0.065 p Value 0.27 0.13 0.37 0.58 0.72 0.25 0.88 0.86 0.29 nCohort Non- 50 52 112 75 73 120 85 86 125 persistent nCohort Persistent 101 98 38 76 77 30 66 64 25 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 75% 77% 79% 74% 74% 83% 74% 77% 84% Specificity 26% 29% 27% 24% 25% 28% 25% 27% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 52% 53% 58% 53% 51% 63% 50% 48% 68% Specificity 54% 56% 53% 52% 51% 53% 49% 49% 54% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 29% 29% 34% 30% 29% 33% 27% 25% 32% Specificity 82% 81% 78% 80% 78% 77% 76% 74% 76% OR Quartile 2 1.07 1.32 1.37 0.884 0.933 1.90 0.946 1.19 1.96 p Value 0.87 0.47 0.48 0.74 0.85 0.23 0.88 0.65 0.25 Lower limit of 0.491 0.618 0.566 0.424 0.446 0.670 0.451 0.563 0.628 95% CI Upper limit of 2.32 2.83 3.32 1.85 1.95 5.37 1.98 2.52 6.13 95% CI OR Quartile 3 1.30 1.43 1.53 1.20 1.05 1.97 0.977 0.897 2.45 p Value 0.45 0.30 0.26 0.57 0.87 0.11 0.94 0.74 0.053 Lower limit of 0.657 0.725 0.728 0.636 0.556 0.865 0.513 0.469 0.987 95% CI Upper limit of 2.56 2.80 3.22 2.28 2.00 4.50 1.86 1.71 6.10 95% CI OR Quartile 4 1.83 1.68 1.81 1.74 1.42 1.64 1.22 0.970 1.49 p Value 0.16 0.21 0.15 0.15 0.35 0.26 0.60 0.94 0.40 Lower limit of 0.792 0.742 0.810 0.822 0.678 0.689 0.583 0.460 0.585 95% CI Upper limit of 4.25 3.80 4.04 3.67 3.00 3.92 2.55 2.04 3.80 95% CI

TABLE 33.7 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the “persistent” and “non-persistent” cohorts where persistence starts within 48 hours after sample collection and renal status is assessed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. sCr or UO Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 330 386 368 376 373 355 Average 443 550 468 556 473 561 Stdev 377 563 366 610 364 635 p (t-test) 0.23 0.29 0.29 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 49 102 70 81 79 72 sCr only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 324 392 348 378 371 374 Average 434 555 453 567 458 576 Stdev 382 566 362 613 360 639 p (t-test) 0.17 0.18 0.16 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 51 99 70 80 79 71 UO only Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort Median 364 392 337 396 334 397 Average 493 540 494 548 500 535 Stdev 540 421 533 420 535 393 p (t-test) 0.60 0.57 0.72 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1510 n (Patient) 109 47 111 39 116 34 Persistence Period Duration (hr) 24 48 72 sCr sCr sCr UO sCr only UO only UO sCr only UO only UO sCr only UO only AUC 0.56 0.58 0.54 0.53 0.55 0.56 0.52 0.54 0.56 SE 0.049 0.048 0.051 0.047 0.047 0.054 0.047 0.047 0.057 p Value 0.22 0.097 0.39 0.58 0.32 0.28 0.69 0.36 0.33 nCohort Non- 49 51 103 70 70 111 79 79 116 persistent nCohort Persistent 102 99 47 81 80 39 72 71 34 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 75% 77% 77% 74% 75% 79% 75% 76% 79% Specificity 27% 29% 26% 24% 26% 27% 25% 27% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 53% 54% 53% 52% 52% 56% 50% 51% 59% Specificity 55% 57% 51% 51% 53% 52% 49% 51% 53% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 28% 28% 34% 30% 30% 33% 28% 28% 32% Specificity 82% 80% 79% 80% 80% 77% 77% 77% 77% OR Quartile 2 1.11 1.38 1.16 0.916 0.916 1.44 1.02 1.15 1.41 p Value 0.79 0.41 0.71 0.82 0.82 0.42 0.96 0.71 0.47 Lower limit of 0.511 0.643 0.520 0.438 0.438 0.594 0.487 0.549 0.556 95% CI Upper limit of 2.42 2.92 2.60 1.92 1.92 3.47 2.12 2.41 3.56 95% CI OR Quartile 3 1.38 1.52 1.20 1.14 1.14 1.42 0.975 1.05 1.58 p Value 0.36 0.23 0.60 0.69 0.69 0.35 0.94 0.87 0.24 Lower limit of 0.697 0.769 0.604 0.601 0.601 0.679 0.515 0.556 0.731 95% CI Upper limit of 2.74 3.00 2.40 2.16 2.16 2.95 1.85 2.00 3.44 95% CI OR Quartile 4 1.77 1.62 1.90 1.68 1.68 1.72 1.30 1.33 1.58 p Value 0.19 0.25 0.10 0.18 0.18 0.18 0.48 0.45 0.29 Lower limit of 0.761 0.714 0.884 0.791 0.791 0.772 0.624 0.636 0.682 95% CI Upper limit of 4.10 3.66 4.09 3.58 3.58 3.83 2.72 2.78 3.64 95% CI

TABLE 33.8 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the persistant and non-persistant cohorts where persistance starts within 72 hours after sample collection and renal status is assesed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 330 386 364 378 364 378 Average 443 550 465 557 463 567 Stdev 377 563 368 606 368 620 p (t-test) 0.23 0.27 0.22 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 49 102 69 82 75 76 sCr only Median 324 392 332 380 332 380 Average 434 555 450 568 450 578 Stdev 382 566 364 610 364 624 p (t-test) 0.17 0.16 0.13 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 51 99 69 81 75 75 UO only Median 334 394 332 396 331 397 Average 491 538 495 538 501 527 Stdev 551 408 544 403 546 376 p (t-test) 0.59 0.63 0.78 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1510 n (Patient) 98 52 105 45 110 40 Persistence Period Duration (hr) 24 24 24 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.56 0.58 0.55 0.53 0.55 0.56 0.54 0.56 0.56 SE 0.049 0.048 0.050 0.047 0.047 0.052 0.047 0.047 0.054 p Value 0.22 0.097 0.28 0.49 0.26 0.25 0.38 0.18 0.30 nCohort Non-persistent 49 51 98 69 69 105 75 75 110 nCohort Persistent 102 99 52 82 81 45 76 75 40 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 75% 77% 77% 74% 75% 80% 76% 77% 80% Specificity 27% 29% 27% 25% 26% 28% 27% 28% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 53% 54% 56% 52% 53% 58% 53% 53% 60% Specificity 55% 57% 53% 52% 54% 53% 52% 53% 54% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 28% 28% 33% 29% 30% 31% 28% 28% 30% Specificity 82% 80% 79% 80% 80% 77% 77% 77% 76% OR Quartile 2 1.11 1.38 1.20 0.950 1.08 1.53 1.17 1.33 1.50 p Value 0.79 0.41 0.64 0.89 0.84 0.33 0.67 0.45 0.37 Lower limit of 95% CI 0.511 0.643 0.549 0.454 0.515 0.655 0.561 0.634 0.621 Upper limit of 95% CI 2.42 2.95 2.64 1.99 2.25 3.56 2.45 2.78 3.62 OR Quartile 3 1.38 1.52 1.43 1.20 1.31 1.56 1.20 1.31 1.74 p Value 0.36 0.23 0.30 0.57 0.41 0.21 0.57 0.41 0.14 Lower limit of 95% CI 0.697 0.769 0.725 0.634 0.687 0.773 0.636 0.688 0.832 Upper limit of 95% CI 2.74 3.00 2.80 2.28 2.49 3.16 2.28 2.48 3.62 OR Quartile 4 1.77 1.62 1.78 1.63 1.65 1.52 1.30 1.33 1.38 p Value 0.19 0.25 0.13 0.21 0.19 0.29 0.48 0.45 0.43 Lower limit of 95% CI 0.761 0.714 0.838 0.764 0.777 0.700 0.623 0.634 0.618 Upper limit of 95% CI 4.10 3.66 3.79 3.46 3.52 3.32 2.73 2.78 3.10

TABLE 33.9 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the persistant and non-persistant cohorts where persistance starts within 96 hours after sample collection and renal status is assesed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 330 386 348 380 348 380 Average 443 550 465 557 463 566 Stdev 377 563 370 602 370 616 p (t-test) 0.23 0.27 0.22 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 49 102 68 83 74 77 sCr only Median 324 392 331 386 331 386 Average 434 555 449 567 449 577 Stdev 382 566 366 606 366 620 p (t-test) 0.17 0.16 0.13 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 51 99 68 82 74 76 UO only Median 330 397 329 412 328 428 Average 483 551 486 550 493 541 Stdev 554 408 551 399 552 375 p (t-test) 0.43 0.47 0.60 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1510 n (Patient) 95 55 100 50 105 45 Persistence Period Duration (hr) 24 48 72 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.56 0.58 0.57 0.54 0.56 0.58 0.55 0.57 0.58 SE 0.049 0.048 0.049 0.047 0.047 0.050 0.047 0.047 0.052 p Value 0.22 0.097 0.13 0.45 0.22 0.11 0.34 0.16 0.14 nCohort Non-persistent 49 51 95 68 68 100 74 74 105 nCohort Persistent 102 99 55 83 82 50 77 76 45 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 75% 77% 78% 75% 76% 80% 77% 78% 80% Specificity 27% 29% 27% 25% 26% 28% 27% 28% 28% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 53% 54% 58% 53% 54% 60% 53% 54% 62% Specificity 55% 57% 55% 53% 54% 55% 53% 54% 55% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 28% 28% 33% 29% 29% 32% 27% 28% 31% Specificity 82% 80% 79% 79% 79% 78% 77% 77% 77% OR Quartile 2 1.11 1.38 1.35 0.984 1.12 1.56 1.21 1.38 1.53 p Value 0.79 0.41 0.45 0.97 0.77 0.29 0.61 0.40 0.33 Lower limit of 95% CI 0.511 0.643 0.617 0.470 0.534 0.686 0.582 0.657 0.655 Upper limit of 95% CI 2.42 2.95 2.95 2.06 2.33 3.53 2.53 2.88 3.56 OR Quartile 3 1.38 1.52 1.68 1.27 1.38 1.83 1.27 1.38 2.03 p Value 0.36 0.23 0.13 0.47 0.33 0.085 0.47 0.33 0.052 Lower limit of 95% CI 0.697 0.769 0.860 0.668 0.725 0.920 0.670 0.725 0.994 Upper limit of 95% CI 2.74 3.00 3.29 2.41 2.63 3.65 2.40 2.62 4.15 OR Quartile 4 1.77 1.62 1.82 1.57 1.60 1.67 1.26 1.28 1.52 p Value 0.19 0.25 0.12 0.24 0.23 0.19 0.54 0.51 0.29 Lower limit of 95% CI 0.761 0.714 0.863 0.737 0.749 0.781 0.601 0.611 0.700 Upper limit of 95% CI 4.10 3.66 3.86 3.34 3.40 3.57 2.63 2.68 3.32

TABLE 33.10 Comparison of marker levels and the area under the ROC curve (AUC) in EDTA samples for the persistant and non-persistant cohorts where persistance starts within168 hours after sample collection and renal status is assesed by serum creatinine (sCr) only, urine output (UO) only, or serum creatinine or urine output RIFLE criteria. Persistence Period Duration (hr) 24 48 72 Non- Non- Non- persistent Persistent persistent Persistent persistent Persistent Cohort Cohort Cohort Cohort Cohort Cohort sCr or UO Median 330 386 330 394 331 392 Average 432 553 442 570 447 574 Stdev 374 561 357 599 358 611 p (t-test) 0.18 0.13 0.13 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 47 104 65 86 70 81 sCr only Median 327 386 331 386 331 386 Average 436 553 450 564 449 573 Stdev 386 564 371 599 371 612 p (t-test) 0.19 0.18 0.14 Min 46.8 50.8 46.8 50.8 46.8 50.8 Max 1880 4690 1880 4690 1880 4690 n (Patient) 50 100 66 84 72 78 UO only Median 329 412 328 428 328 428 Average 484 545 484 548 495 533 Stdev 563 399 558 400 561 378 p (t-test) 0.48 0.46 0.67 Min 50.8 46.8 50.8 46.8 50.8 46.8 Max 4690 1700 4690 1700 4690 1510 n (Patient) 92 58 95 55 99 51 Persistence Period Duration (hr) 24 48 72 sCr sCr UO sCr sCr UO sCr sCr UO or UO only only or UO only only or UO only only AUC 0.57 0.58 0.58 0.56 0.56 0.58 0.56 0.57 0.57 SE 0.049 0.049 0.048 0.047 0.047 0.049 0.047 0.047 0.050 p Value 0.14 0.10 0.11 0.18 0.20 0.10 0.20 0.14 0.18 nCohort Non-persistent 47 50 92 65 66 95 70 72 99 nCohort Persistent 104 100 58 86 84 55 81 78 51 Cutoff Quartile 2 218 217 217 218 217 217 218 217 217 Sensitivity 76% 77% 79% 76% 76% 80% 77% 78% 78% Specificity 28% 30% 28% 26% 27% 28% 27% 29% 27% Cutoff Quartile 3 373 372 372 373 372 372 373 372 372 Sensitivity 53% 53% 59% 55% 54% 60% 54% 54% 61% Specificity 55% 56% 55% 55% 55% 56% 54% 54% 56% Cutoff Quartile 4 674 661 661 674 661 661 674 661 661 Sensitivity 29% 28% 31% 30% 29% 31% 28% 27% 29% Specificity 83% 80% 78% 82% 79% 78% 79% 76% 77% OR Quartile 2 1.21 1.43 1.51 1.10 1.20 1.59 1.22 1.48 1.36 p Value 0.64 0.35 0.30 0.81 0.63 0.26 0.60 0.30 0.45 Lower limit of 95% CI 0.553 0.669 0.692 0.523 0.573 0.716 0.582 0.705 0.612 Upper limit of 95% CI 2.64 3.08 3.30 2.30 2.51 3.52 2.54 3.10 3.04 OR Quartile 3 1.39 1.44 1.76 1.50 1.38 1.89 1.41 1.38 1.94 p Value 0.35 0.30 0.095 0.22 0.32 0.064 0.29 0.33 0.059 Lower limit of 95% CI 0.696 0.725 0.906 0.783 0.725 0.964 0.743 0.725 0.974 Upper limit of 95% CI 2.78 2.84 3.43 2.86 2.64 3.72 2.68 2.62 3.85 OR Quartile 4 1.98 1.56 1.62 1.91 1.49 1.58 1.45 1.19 1.38 p Value 0.13 0.29 0.20 0.10 0.31 0.23 0.33 0.64 0.41 Lower limit of 95% CI 0.827 0.686 0.769 0.880 0.697 0.745 0.688 0.569 0.643 Upper limit of 95% CI 4.72 3.53 3.41 4.16 3.17 3.34 3.07 2.50 2.95

While the invention has been described and exemplified in sufficient detail for those skilled in this art to make and use it, various alternatives, modifications, and improvements should be apparent without departing from the spirit and scope of the invention. The examples provided herein are representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. Modifications therein and other uses will occur to those skilled in the art. These modifications are encompassed within the spirit of the invention and are defined by the scope of the claims.

It will be readily apparent to a person skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention.

All patents and publications mentioned in the specification are indicative of the levels of those of ordinary skill in the art to which the invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

The invention illustratively described herein suitably may be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of” and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims.

Other embodiments are set forth within the following claims. 

1-111. (canceled)
 112. A method of treating a subject having acute kidney injury, the method comprising: (a) detecting a level of Metalloproteinase inhibitor 2 in a body fluid sample obtained from the subject; (b) determining the subject is at an increased risk of not recovering from acute kidney injury when the detected level of Metalloproteinase inhibitor 2 is above a predetermined threshold level of Metalloproteinase inhibitor 2, wherein recovery is a maximum RIFLE stage of RIFLE stage I during a period no less than 24 hours, the period beginning within 7 days after sample collection; and (c) treating the subject based on the subject's increased risk of not recovering from acute kidney injury for persistent acute kidney injury, wherein the treatment comprises renal replacement therapy, withdrawing delivery of compounds that are known to be damaging to the kidney, delaying procedures that are known to be damaging to the kidney, or modifying diuretic administration.
 113. The method of claim 112, wherein the period is no less than 48 hours.
 114. The method of claim 112, wherein the period is no less than 72 hours.
 115. The method of claim 112, wherein the period begins within 96 hours after sample collection.
 116. The method of claim 112, wherein the period begins within 72 hours after sample collection.
 117. The method of claim 112, wherein the period begins within 48 hours after sample collection.
 118. The method of claim 112, wherein the body fluid sample is a urine sample.
 119. The method of claim 118, wherein the period is no less than 72 hours and begins within 48 hours after sample collection.
 120. The method of claim 112, wherein recovery is a maximum RIFLE stage of RIFLE stage R during the period.
 121. The method of claim 120, wherein the subject has RIFLE stage F acute kidney injury at the time of sample collection.
 122. The method of claim 120, wherein the subject has RIFLE stage I acute kidney injury at the time of sample collection.
 123. The method of claim 120, wherein the period is no less than 48 hours.
 124. The method of claim 120, wherein the period is no less than 72 hours.
 125. The method of claim 120, wherein the period begins within 96 hours after sample collection.
 126. The method of claim 120, wherein the period begins within 72 hours after sample collection.
 127. The method of claim 120, wherein the period begins within 48 hours after sample collection.
 128. The method of claim 112, wherein recovery is a maximum RIFLE stage of RIFLE stage 0 during the period.
 129. The method of claim 128, wherein the subject R stage I or F acute kidney injury at the time of sample collection.
 130. The method of claim 128, wherein the period is no less than 48 hours.
 131. The method of claim 128, wherein the period is no less than 72 hours.
 132. The method of claim 128, wherein the period begins within 96 hours after sample collection.
 133. The method of claim 128, wherein the period begins within 72 hours after sample collection.
 134. The method of claim 128, wherein the period begins within 48 hours after sample collection. 